Pharmaceutical combinations comprising a bispecific antibody and an oncolytic virus and uses thereof

Abstract

This invention belongs to the fields of tumor therapy and molecular immunology, and relates to a drug combination comprising a bispecific antibody and an oncolytic virus, and its uses. Specifically, it relates to a drug combination comprising an anti-PD-1-anti-VEGFA bispecific antibody and an oncolytic virus, and its uses. The drug combination of this invention can effectively treat or prevent tumors and has good application prospects.

Description

[0001] Cross-references to related applications

[0002] This application is based on and claims priority to CN application number 202510466145.9, filed on April 14, 2025, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the fields of tumor therapy and molecular immunology, and relates to a drug combination comprising a bispecific antibody and an oncolytic virus, and its use. Specifically, it relates to a drug combination comprising an anti-PD-1-anti-VEGFA bispecific antibody and an oncolytic virus, and its use. Background Technology

[0004] Cancer, especially malignant cancer, is a serious threat to human health worldwide, ranking second among all causes of death. Moreover, its incidence has shown a significant upward trend in recent years. Malignant cancers have poor treatment outcomes, high rates of late-stage metastasis, and generally poor prognoses. While current clinical treatments such as radiotherapy, chemotherapy, and surgery have significantly alleviated pain and prolonged survival, these methods have considerable limitations, making further improvements in efficacy difficult.

[0005] Tumor growth has two distinct phases: a slow, avascular growth phase followed by a rapid, vascularized proliferation phase. Angiogenesis allows the tumor to obtain sufficient nutrients to complete the angiogenesis transition. Without angiogenesis, the primary tumor grows to no more than 1-2 mm, and metastasis cannot occur.

[0006] Vascular endothelial growth factor (VEGF) is a class of growth factors that promote endothelial cell proliferation, angiogenesis, and vascular permeability. It binds to VEGF receptors on the cell surface and functions by activating tyrosine kinase signaling pathways. In tumor tissues, tumor cells, invading macrophages, and mast cells secrete high levels of VEGF, which stimulates tumor vascular endothelial cells in a paracrine manner, promoting endothelial cell proliferation and migration, inducing angiogenesis, promoting sustained tumor growth, increasing vascular permeability, causing fibrin deposition in surrounding tissues, promoting the infiltration of monocytes, fibroblasts, and endothelial cells, facilitating tumor matrix formation and tumor cell entry into new blood vessels, and promoting tumor metastasis. Therefore, inhibiting tumor angiogenesis is considered one of the most promising cancer treatment methods. The VEGF family includes: VEGFA, VEGFB, VEGFC, VEGRD, and PIGF. Vascular endothelial growth factor receptors (VEGFRs) include VEGFR1 (also known as Flt1), VEGFR2 (also known as KDR or Flk1), VEGFR3 (also known as Flt4), and Neuropilin-1 (NRP-1). The first three receptors are structurally similar, all belonging to the tyrosine kinase superfamily, and each consists of an extramembrane domain, a transmembrane segment, and an intramembrane domain. The extramembrane domain is composed of an immunoglobulin-like domain, while the intramembrane domain is a tyrosine kinase domain. VEGFR1 and VEGFR2 are mainly located on the surface of vascular endothelial cells, while VEGFR3 is mainly located on the surface of lymphatic endothelial cells.

[0007] VEGF family molecules have different affinities for several receptors. VEGFA mainly binds to VEGFR1, VEGFR2 and NRP-1 to exert its function. VEGFR1 was the first receptor to be discovered. Under normal physiological conditions, VEGFR1 has a higher affinity for VEGFA than VEGFR2, but its intracellular tyrosinase activity is lower than that of VEGFR2 (Ma Li, Chinese Journal of Eugenics and Genetics, 24(5)(2016): 146-148).

[0008] VEGFR2 is a major regulator of angiogenesis and angiogenesis, and it has a higher tyrosine kinase activity than VEGFR1. After VEGFR2 binds to its ligand VEGFA, it mediates the proliferation and differentiation of vascular endothelial cells, as well as the process of angiogenesis and vascular permeability (Roskoski R Jr. et al., Crit Rev Oncol Hematol, 62(3)(2007):179-213.). After VEGFA binds to VEGFR2, it mediates the transcriptional expression of intracellular related protein genes through the downstream PLC-γ-PKC-Raf-MEK-MAPK signaling pathway, promoting the proliferation of vascular endothelial cells (Takahashi T et al., Oncogene, 18(13)(1999):2221-2230.).

[0009] VEGFR3 belongs to the tyrosine kinase family and is mainly expressed in vascular endothelial cells during embryonic development and lymphatic endothelial cells during adulthood. VEGFC and VEGGFD bind to VEGFR3 to stimulate the proliferation and migration of lymphatic endothelial cells, promoting the formation of lymphatic vessels. NRP-1 is a non-tyrosine kinase transmembrane protein that cannot independently transduce biological signals. It can only mediate signal transduction after forming a complex with the VEGF tyrosine kinase receptor. (Ma Li, Chinese Journal of Eugenics and Genetics, 24(5)(2016): 146-148).

[0010] VEGFA and VEGFR2 are mainly involved in regulating angiogenesis. Before and after VEGFA and VEGFR2 bind, they trigger a series of intermediate signals in the upstream and downstream pathways to form a cascade reaction, which ultimately changes the physiological functions of endothelial cells in different forms such as proliferation, survival, migration, increased permeability and infiltration into surrounding tissues (Dong Hongchao et al., Modern Oncology Medicine, Vol. 22, No. 9, September 2014, pp. 2231-3).

[0011] Programmed cell death protein-1 (PD-1), also known as CD279, is a type I transmembrane glycoprotein receptor belonging to the CD28 immunoglobulin superfamily. It is widely expressed on T cells, B cells, and myeloid cells. PD-1 has two natural ligands, PD-L1 and PD-L2. Both PD-L1 and PD-L2 belong to the B7 superfamily and are constitutively or inducibly expressed on the surface of various cell membranes, including non-hematopoietic cells and various tumor cells. PD-L1 is mainly expressed on T cells, B cells, dendritic cells (DCs), microvascular endothelial cells, and various tumor cells. PD-L2 is expressed only on antigen-presenting cells such as dendritic cells and macrophages. The interaction between PD-1 and its ligands can inhibit lymphocyte activation, suppress T cell proliferation, and inhibit the secretion of cytokines such as IL-2 and IFN-γ.

[0012] Numerous studies have shown that the tumor microenvironment protects tumor cells from damage by immune cells. PD-1 expression is upregulated in lymphocytes infiltrating the tumor microenvironment, and various primary tumor tissues, such as lung cancer, liver cancer, ovarian cancer, skin cancer, colon cancer, and glioma, show PD-L1 positivity in immunohistochemical analysis. Furthermore, PD-L1 expression in tumors is significantly associated with poor prognosis in cancer patients. Blocking the interaction between PD-1 and its ligands can promote tumor-specific T-cell immunity and improve the efficiency of tumor cell immune clearance. Numerous clinical trials have demonstrated that antibodies targeting PD-1 or PD-L1 can promote CD8+ T-cell infiltration into tumor tissues and upregulate anti-tumor immune effector factors such as IL-2, IFN-γ, granzyme B, and perforin, thereby effectively inhibiting tumor growth.

[0013] In addition, anti-PD-1 antibodies can also be used to treat chronic viral infections. Chronic viral infections are often accompanied by the loss of function and a reduction in the number of virus-specific effector T cells. By injecting PD-1 antibodies, the interaction between PD-1 and PD-L1 can be blocked, thereby effectively inhibiting the depletion of effector T cells in chronic viral infections.

[0014] Bispecific antibodies, also known as bispecific antibodies, are specific drugs that simultaneously target two different antigens. They can be produced through immunosorting and purification. Alternatively, they can be obtained through genetic engineering. Genetic engineering offers advantages in terms of flexibility in binding site optimization, synthetic formulation, and yield. Currently, more than 45 forms have been identified (Müller D, Kontermann RE. Bispecific antibodies for cancer immunotherapy: Current perspectives. BioDrugs 2010; 24:89-98). Currently, many bispecific antibodies have been developed in the form of IgG-ScFv, also known as the Morrison model (Coloma MJ, Morrison SL Design and production of novel tetravalent bispecific antibodies. Nat Biotechnol., 1997;15:159-163). Due to its similarity to naturally occurring IgG, this form has advantages in antibody engineering, expression, and purification, and has been proven to be an ideal form for bispecific antibodies (Miller BR, Demarest SJ, et al., Stability engineering of scFvs for the development of bispecific and multivalent antibodies. Protein Eng DesSel 2010; 23:549-57; Fitzgerald J, Lugovskoy A. Rational engineering of antibody therapeutics targeting multiple oncogene pathways. MAbs 2011; 3:299-309). Ivonescimab is the world's first approved bispecific antibody drug with a "tumor immunity + anti-angiogenesis" mechanism, capable of simultaneously targeting PD-1 and VEGF. Ivonescimab has already accumulated substantial high-level evidence of its efficacy and safety in lung cancer.

[0015] Oncolytic virus (OV) therapy has been considered a promising new treatment for cancer. It is a treatment regimen that specifically destroys cancer cells through the oncolytic effect. Its killing mechanism is characterized by the lysis of cancer cells through viral replication, which induces strong tumor lysis and induces systemic tumor-specific immunity, while causing significantly fewer side effects than chemotherapy or radiotherapy.

[0016] Among various oncolytic viruses, herpes simplex virus type 1 (“HSV-1”)-based OVs are the most advanced. For example, the herpes simplex virus-based OV (T-Vec) has been approved by the U.S. FDA for the treatment of melanoma. Representative examples of HSV vectors include those described in U.S. Patent Nos. 7,223,593, 7,537,924, 7,063,835, 7,063,851, 7,118,755, 8,277,818, and 8,680,068.

[0017] One challenge with oncolytic herpesvirus vectors is the neurotropic nature of HSV. The viral protein ICP34.5 is involved in viral neurotoxicity, leading to a common strategy of deleting ICP34.5 from vectors used in oncolytic virus therapy. However, complete deletion of ICP34.5 reduces the virus's ability to replicate in tumor tissue by approximately 10-fold. Recombinant vectors, on the other hand, are controlled at both the transcriptional and post-transcriptional (translational) levels to provide more precise control over the virus's oncolytic potential.

[0018] More effective technologies for treating tumors are still needed. Summary of the Invention

[0019] Through in-depth research and creative work, the inventors have innovatively combined an anti-PD-1-anti-VEGFA bispecific antibody with oncolytic virus VG2025. The inventors have surprisingly discovered that this combination achieves effective inhibitory activity against tumor growth, and the effect is superior to either the anti-PD-1-anti-VEGFA bispecific antibody alone or the oncolytic virus VG2025 alone. Therefore, the following invention is provided: One aspect of the present invention relates to a pharmaceutical combination comprising an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody and an oncolytic virus, wherein: The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; Wherein, the first protein functional region is an immunoglobulin and the second protein functional region is a single-chain antibody; or, the first protein functional region is a single-chain antibody and the second protein functional region is an immunoglobulin. in, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; or, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; And, the oncolytic virus is herpes simplex virus type 1 (HSV-1).

[0020] In some embodiments of the present invention, the drug combination, wherein... The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 14; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. or, The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 16; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 14.

[0021] In some embodiments of the invention, the drug combination wherein the heavy chain constant region of the immunoglobulin adopts the Ig gamma-1 chain C region and the light chain constant region adopts the Ig kappa chain C region.

[0022] In some embodiments of the invention, the drug combination wherein the immunoglobulin is human IgG1 subtype; preferably, according to the EU numbering system, the heavy chain constant region of the immunoglobulin has one of the following mutation combinations: L234A and L235A; L234A and G237A; L235A and G237A; or L234A, L235A and G237A.

[0023] In some embodiments of the present invention, the drug combination wherein the amino acid sequence of the heavy chain of the immunoglobulin is as shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is as shown in SEQ ID NO: 18.

[0024] In some embodiments of the present invention, the drug combination wherein the first protein functional region is directly connected to the second protein functional region or connected through a first linker; and / or the heavy chain variable region of the single-chain antibody is directly connected to the light chain variable region of the single-chain antibody or connected through a second linker; Preferably, the first linker and the second linker are independently polypeptides consisting of n peptides shown in SEQ ID NO: 19 connected in series, where n is a positive integer; preferably, n is 1, 2, 3, 4, 5 or 6. Preferably, the first linker and the second linker are independently polypeptides as shown in SEQ ID NO: 20.

[0025] In some embodiments of the present invention, the drug combination, wherein... The single-chain antibody is independently connected in sequence as follows: heavy chain variable region-linker-light chain variable region (VH-linker-VL), or light chain variable region-linker-heavy chain variable region (VL-linker-VH). Preferably, the amino acid sequence of the single-chain antibody is shown in SEQ ID NO: 21.

[0026] In some embodiments of the present invention, the drug combination, wherein... The first protein functional region and the second protein functional region are independently one, two, or more. Preferably, the single-chain antibody is linked to the C-terminus or N-terminus of the heavy chain of an immunoglobulin; Preferably, the bispecific antibody is a bispecific antibody in the form of IgG-scFv; Preferably, the bispecific antibody is a tetravalent bispecific antibody in the form of IgG-scFv.

[0027] In some embodiments of the invention, the drug combination includes a single-chain antibody linked to the C-terminus of the heavy chain of an immunoglobulin. Since immunoglobulins consist of two heavy chains, one immunoglobulin molecule is linked to two single-chain antibody molecules. Preferably, the two single-chain antibody molecules are identical.

[0028] In some embodiments of the present invention, the drug combination includes two single-chain antibodies, one end of each single-chain antibody being attached to the C-terminus or N-terminus of the two heavy chains of an immunoglobulin.

[0029] In some embodiments of the present invention, the V of the single-chain antibody H With V L Disulfide bonds exist between them. In the antibody's V... H and V L The method of introducing disulfide bonds between them is well known in the art, and can be found, for example, in US Patent 5,747,654; Rajagopal et al, Prot. Engin. 10 (1997) 1453-1459; Reiter et al, Nat. Biotechnol. 14 (1996) 1239-1245; Reiter et al, Protein Engineering 8 (1995) 1323-1331; Webber et al, Molecular Immunology 32 (1995) 249-258; Reiter et al, Immunity 2 (1995) 281-287; Reiter et al, JBC 269 (1994) 18327-18331; Reiter et al, Inter. J. of Cancer 58 (1994) 142-149; or Reiter et al., Cancer Res. 54 (1994) 2714-2718; which is incorporated herein by reference.

[0030] In some embodiments of the present invention, the drug combination, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of the first linker and the second linker are both as shown in SEQ ID NO: 20.

[0031] In some embodiments of the present invention, the drug combination, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of the first linker and the second linker are both as shown in SEQ ID NO: 20.

[0032] In some embodiments of the present invention, the drug combination, wherein the anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains (the heavy chain of the anti-PD-1-anti-VEGFA bispecific antibody) and two identical second peptide chains (the light chain of the anti-PD-1-anti-VEGFA bispecific antibody), wherein, The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; Preferably, the first peptide chain and the second peptide chain, and / or the two first peptide chains are independently connected by one or more (e.g., 2, 3 or 4) disulfide bonds; Preferably, there are no disulfide bonds connecting the two second peptide chains.

[0033] In some embodiments of the present invention, the drug combination is wherein the oncolytic virus is a modified HSV-1, preferably VG2025.

[0034] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified (including but not limited to: recombinant, modified) wild-type HSV-1, preferably a modified wild-type HSV-1 strain 17+.

[0035] In some embodiments of the invention, the drug combination wherein the oncolytic virus ICP34.5 is present at a lower level in normal cells (e.g., normal nerve cells) than in tumor cells.

[0036] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified HSV-1, and relative to wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the second copy is downregulated at the posttranscriptional mRNA level; Preferably, the 3'UTR region of the second copy of the ICP34.5 gene contains one or more (e.g., 2-10, preferably 5) miRNA target sequences; Preferably, the miRNA target sequence targets miR-124 and / or miR-143; Preferably, the sequence of miR-124 is as shown in SEQ ID NO: 30; Preferably, the sequence of miR-143 is shown in SEQ ID NO: 31.

[0037] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified HSV-1, and the natural promoter of ICP27 is replaced by a tumor-specific promoter; preferably, replaced by a human carcinoembryonic antigen (CEA) promoter; preferably, the sequence of the CEA promoter is shown in SEQ ID NO: 23.

[0038] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified HSV-1 and comprises an expression cassette expressing one or more of IL12, IL15 and the IL15 receptor α subunit (IL-15Rα); Preferably, the amino acid sequence of IL12 is shown in SEQ ID NO: 26; Preferably, the amino acid sequence of IL15 is shown in SEQ ID NO: 27; Preferably, the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; Preferably, the expression cassette is driven by the CXCR4 promoter; preferably, the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25; Preferably, IL-12, IL-15 and IL-15Rα are tandemly connected, and the N-terminus of IL-15 and IL-15Rα are respectively linked to the 2A self-cleaving peptide; preferably, the amino acid sequence of the 2A self-cleaving peptide is as shown in SEQ ID NO: 28.

[0039] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified HSV-1, and the C-terminal amino acid positions 877-904 of glycoprotein B (gB) are truncated; preferably, the amino acid sequence of the truncated glycoprotein B is shown in SEQ ID NO: 24.

[0040] In some embodiments of the present invention, the drug combination wherein the oncolytic virus is a modified wild-type HSV-1 or a modified wild-type HSV-1 strain 17+, and has any one, any two, any three, or all four of the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24.

[0041] In some embodiments of the present invention, the drug combination, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24.

[0042] In some embodiments of the present invention, the drug combination, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24.

[0043] In some embodiments of the present invention, the drug combination, wherein... The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, as well as the two first peptide chains, are independently connected by one or more disulfide bonds. The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24.

[0044] In some embodiments of the present invention, the drug combination, wherein the anti-PD-1-anti-VEGFA bispecific antibody is mixed with an oncolytic virus, is a pharmaceutical composition; or

[0045] The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are packaged in separate containers, and the drug combination is a combination product.

[0046] In some embodiments of the present invention, the drug combination, wherein the unit dose of the anti-PD-1-anti-VEGFA bispecific antibody is calculated according to the mass of the anti-PD-1-anti-VEGFA bispecific antibody as 100mg-2500mg, 100mg-2000mg, 100mg-1500mg, 100mg-1200mg, 100mg-1000mg, 200mg-800mg, 200mg-500mg, 300mg-600mg, 400mg-500mg, or 450mg.

[0047] In some embodiments of the present invention, the drug combination contains an oncolytic virus titer of approximately 5 × 10⁻⁶. 4 pfu / mL - approximately 2 × 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 10 7 pfu / mL, approximately 10 7 pfu / mL - approximately 10 8 pfu / mL, approximately 10 8 pfu / mL - approximately 10 9 pfu / mL or approximately 10 8 pfu / mL.

[0048] In some embodiments of the present invention, the pharmaceutical combination further comprises one or more pharmaceutically acceptable excipients; preferably, it further comprises a product instruction manual.

[0049] In some embodiments of the present invention, the drug combination comprises the active pharmaceutical ingredient (API) or active ingredient consisting of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus.

[0050] In some embodiments of the invention, the drug combination comprises evasin and VG2025.

[0051] In some embodiments of the present invention, the drug combination further comprises an effective amount of a chemotherapy drug, wherein the chemotherapy drug is a chemotherapy drug of the mFOLFOX6 regimen, the FOLFIRI regimen, or the CapeOX regimen.

[0052] In some embodiments of the present invention, the drug combination comprises the anti-PD-1-anti-VEGFA bispecific antibody, the oncolytic virus, and a chemotherapeutic drug, wherein the chemotherapeutic drug is a chemotherapeutic drug of the mFOLFOX6 regimen, the FOLFIRI regimen, or the CapeOX regimen.

[0053] In some embodiments of the present invention, the drug combination comprises an active pharmaceutical ingredient (API) of ivoxil, VG2025, and a chemotherapy drug, wherein the chemotherapy drug is a chemotherapy drug of the mFOLFOX6 regimen, the FOLFIRI regimen, or the CapeOX regimen.

[0054] In some embodiments of the present invention, the drug combination further comprises an effective amount of a chemotherapy drug, wherein the chemotherapy drug is oxaliplatin, leucovorin (LV), and 5-fluorouracil (5-FU).

[0055] In some embodiments of the present invention, the drug combination further comprises an effective amount of a chemotherapy drug, wherein the chemotherapy drug is irinotecan, leucovorin calcium, and 5-fluorouracil.

[0056] In some embodiments of the present invention, the drug combination further comprises an effective amount of a chemotherapy drug, wherein the chemotherapy drug is oxaliplatin and capecitabine.

[0057] In some embodiments of the present invention, the drug combination comprises the pharmacodynamic ingredient (API) or active ingredient consisting of the anti-PD-1-anti-VEGFA bispecific antibody, the oncolytic virus, and a chemotherapeutic drug, wherein the chemotherapeutic drug is oxaliplatin, leucovorin calcium, and 5-fluorouracil.

[0058] In some embodiments of the present invention, the drug combination comprises the pharmacodynamic ingredient (API) or active ingredient consisting of the anti-PD-1-anti-VEGFA bispecific antibody, the oncolytic virus, and a chemotherapeutic drug, wherein the chemotherapeutic drug is irinotecan, leucovorin calcium, and 5-fluorouracil.

[0059] In some embodiments of the present invention, the drug combination comprises the pharmacodynamic ingredient (API) or active ingredient consisting of the anti-PD-1-anti-VEGFA bispecific antibody, the oncolytic virus, and a chemotherapeutic drug, wherein the chemotherapeutic drug is oxaliplatin and capecitabine.

[0060] In some embodiments of the present invention, the drug combination comprises an active pharmaceutical ingredient (API) of ivoxil, VG2025, and a chemotherapy drug, wherein the chemotherapy drug is oxaliplatin, leucovorin calcium, and 5-fluorouracil.

[0061] In some embodiments of the present invention, the drug combination comprises an active pharmaceutical ingredient (API) of ivoxil, VG2025, and a chemotherapy drug, wherein the chemotherapy drug is irinotecan, leucovorin calcium, and 5-fluorouracil.

[0062] In some embodiments of the present invention, the drug combination comprises an active pharmaceutical ingredient (API) of ivermectin, VG2025, and a chemotherapy drug, wherein the chemotherapy drug is oxaliplatin and capecitabine.

[0063] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are those used in the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen.

[0064] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are those used in the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen.

[0065] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, as well as the two first peptide chains, are independently connected by one or more disulfide bonds. The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are those used in the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen.

[0066] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapy drug.

[0067] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are oxaliplatin, leucovorin calcium, and 5-fluorouracil.

[0068] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are oxaliplatin, leucovorin calcium, and 5-fluorouracil.

[0069] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, as well as the two first peptide chains, are independently connected by one or more disulfide bonds. The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are oxaliplatin, leucovorin calcium, and 5-fluorouracil.

[0070] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are irinotecan, leucovorin calcium, and 5-fluorouracil.

[0071] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are irinotecan, leucovorin calcium, and 5-fluorouracil.

[0072] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, as well as the two first peptide chains, are independently connected by one or more disulfide bonds. The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are irinotecan, leucovorin calcium, and 5-fluorouracil.

[0073] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs mentioned are oxaliplatin and capecitabine.

[0074] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; Preferably, the amino acid sequences of both the first linker and the second linker are as shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs mentioned are oxaliplatin and capecitabine.

[0075] In some embodiments of the present invention, the drug combination comprises an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus, and a chemotherapeutic drug, wherein... The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, as well as the two first peptide chains, are independently connected by one or more disulfide bonds. The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (1) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (2) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (3) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein, The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (4) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs mentioned are oxaliplatin and capecitabine.

[0076] In some embodiments of the present invention, the drug combination, wherein the anti-PD-1-anti-VEGFA bispecific antibody, the oncolytic virus, and the chemotherapy drug are in separate packages, is a combination product.

[0077] In some embodiments of the present invention, the drug combination is used to treat or prevent tumors; Preferably, the tumor is selected from one or more of colorectal cancer and liver cancer; Preferably, the tumor is an advanced tumor or a metastatic tumor; Preferably, the tumor is a colorectal cancer with liver metastasis; Preferably, the tumor is a late-stage colorectal cancer with liver metastasis; Preferably, the liver cancer is hepatocellular carcinoma, advanced liver cancer, or advanced hepatocellular carcinoma.

[0078] In some embodiments of the present invention, the drug combination is characterized by any one, any two, any three, any four, any five, any six, any seven, or all eight of the following: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Preferably, the anti-PD-1-anti-VEGFA bispecific antibody is administered after the oncolytic virus, and the interval is 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg, 1200 mg, 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg or 200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, or administration once on the first and third days of each cycle, and no administration is given during the rest of the cycle; and each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, oral administration or other cavity perfusion administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen.

[0079] In some embodiments of the present invention, the drug combination, wherein: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 1.0 × 10⁻⁶ per subject. 6 PFU up to 1.0×10 9 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, at a dose of 10-30 mg per kilogram of body weight (e.g., 20 mg), by intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0080] In some embodiments of the present invention, the drug combination, wherein: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once each on days 1 and 3, at a dose of 2.0 × 10⁻⁶ per subject.7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0081] In some embodiments of the present invention, the drug combination, wherein: VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0082] Each pharmaceutical formulation of the drug combination of the present invention can be administered simultaneously (i.e., in the same pharmaceutical composition), in parallel (i.e., one after another in any order as separate pharmaceutical formulations), or sequentially in any order. Sequential administration is particularly useful when the pharmaceutical formulations in the drug combination may be in different dosage forms (one drug is a tablet or capsule and the other is a sterile liquid formulation) and / or at different dosing schedules (e.g., chemotherapeutic agents are administered at least daily and biological agents are administered less frequently (e.g., once a week, once every two weeks, or once every three weeks)).

[0083] Each pharmaceutical preparation in the drug combination of the present invention can be administered orally or parenterally, including intratumoral, intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, local and percutaneous routes.

[0084] In one or more embodiments of the present invention, the pharmaceutical combination is a fixed combination, for example, in the form of a solid pharmaceutical composition or a liquid pharmaceutical composition; or

[0085] The drug combination is not a fixed combination; for example, the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus VG2025 are each in the form of a drug composition.

[0086] Another aspect of the present invention relates to a pillbox product comprising the drug combination described in any one of the present invention, and product instructions.

[0087] In some embodiments of the present invention, the drug combination, wherein... The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is independently 0.1-100 mg per kilogram of body weight, preferably 5-50 mg or 5-25 mg per kilogram of body weight; The anti-PD-1-anti-VEGFA bispecific antibody is administered independently once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks or 3 weeks; and / or The administration methods include intravenous infusion, intravenous injection, subcutaneous injection, or intraperitoneal injection.

[0088] In some embodiments of the present invention, the drug combination, wherein... The single dose of the oncolytic virus VG2025 is approximately 5 × 10⁻⁶. 4 pfu to approximately 2 × 10 9 A range of PFUs is provided for intravenous delivery in humans. For intratumoral injection, a preferred dose range may be approximately 10 ppm per dose. 6 pfu to approximately 10 9 PFU (injectable volume ranging from about 0.1 mL to about 5 mL, preferably administered to the subject in doses of up to 5 mL, up to 4 mL, up to 3 mL, up to 2 mL, up to 1 mL, up to 0.5 mL, up to 0.2 mL, or up to 0.1 mL). In other embodiments, the dose range may be about 10 mL. 6 pfu to approximately 10 7 PFU, or about 10 7 pfu to approximately 10 8 PFU, or about 10 8 pfu to approximately 10 9 PFU can be administered as a single dose or as multiple doses over time. As used herein, PFU refers to "plaque-forming unit".

[0089] In some embodiments of the invention, lower or higher doses than the standard may be used. Therefore, in some embodiments, less than about 10 [units of measurement] may be administered to the patient. 6pfu or more than about 10 9 pfu.

[0090] The oncolytic virus VG2025 is administered periodically. At the beginning of each cycle, it is administered once a day for two or three consecutive days, and no administration is given for the rest of the cycle. After that, it is administered in cycles of two weeks, four weeks, or eight weeks. and / or The administration method can be intratumoral injection, oral administration, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, or other cavity perfusion administration. Preferably, the administration method is intratumoral injection.

[0091] Another aspect of the present invention relates to the use of the pharmaceutical combination described in any one of the present invention in the preparation of a medicament for treating or preventing tumors; Preferably, the tumor is selected from one or more of colorectal cancer and liver cancer; Preferably, the tumor is an advanced tumor or a metastatic tumor; Preferably, the tumor is a colorectal cancer with liver metastasis; Preferably, the tumor is a late-stage colorectal cancer with liver metastasis; Preferably, the liver cancer is hepatocellular carcinoma, advanced liver cancer, or advanced hepatocellular carcinoma.

[0092] In some embodiments of the present invention, the described use is characterized by any one, any two, any three, any four, any five, any six, any seven, or all eight of the following: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Preferably, the anti-PD-1-anti-VEGFA bispecific antibody is administered after the oncolytic virus, and the interval is 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg, 1200 mg, 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg or 200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, and no administration is given during the rest of the cycle. Each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, oral administration or other cavity perfusion administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen.

[0093] In some embodiments of the present invention, the described uses include: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 1.0 × 10⁻⁶ per subject. 6 PFU up to 1.0×10 9 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 10-30 mg per kilogram of body weight (e.g., 20 mg), by intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0094] In some embodiments of the present invention, the described uses include: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once each on days 1 and 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0095] In some embodiments of the present invention, the described uses include: VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0096] Another aspect of the invention relates to a method of treating or preventing tumors, comprising the step of administering to a subject in need an effective amount of any of the drug combinations of the present invention; Preferably, the tumor is selected from one or more of colorectal cancer and liver cancer; Preferably, the tumor is an advanced tumor or a metastatic tumor; Preferably, the tumor is a colorectal cancer with liver metastasis; Preferably, the tumor is a late-stage colorectal cancer with liver metastasis; Preferably, the liver cancer is hepatocellular carcinoma, advanced liver cancer, or advanced hepatocellular carcinoma.

[0097] In some embodiments of the present invention, the method for treating or preventing tumors is characterized by any one, two, three, four, five, six, seven, or all eight of the following: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Preferably, the anti-PD-1-anti-VEGFA bispecific antibody is administered after the oncolytic virus, and the interval is 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg, 1200 mg, 10-1000 mg, 50-500 mg, 100-400 mg, 150-300 mg, 150-250 mg or 200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, and no administration is given during the rest of the cycle. Each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection, oral administration or other cavity perfusion administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen.

[0098] In some embodiments of the present invention, the method for treating or preventing tumors includes: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 1.0 × 10⁻⁶ per subject. 6 PFU up to 1.0×10 9 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 10-30 mg per kilogram of body weight (e.g., 20 mg), by intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0099] In some embodiments of the present invention, the method for treating or preventing tumors includes: The oncolytic virus was administered once daily on days 1-2 or 1-3, or once each on days 1 and 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0100] In some embodiments of the present invention, the method for treating or preventing tumors includes: VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. Preferably, the chemotherapy drugs are administered once on day 8 and once on day 22, according to the mFOLFOX6 regimen, FOLFIRI regimen or CapeOX regimen, by intravenous infusion.

[0101] In some embodiments, the anti-PD-1-anti-VEGFA bispecific antibody or its antigen-binding fragment is administered by injection. Prior to injection, the anti-PD-1-anti-VEGFA bispecific antibody or its antigen-binding fragment needs to be formulated into an injectable form. The preferred injectable form is an injection solution containing the anti-PD-1-anti-VEGFA bispecific antibody or its antigen-binding fragment, and optionally buffers, stabilizers, pH adjusters, and surfactants; preferably, it contains antibody evodiamide. Excipients: histidine, histidine hydrochloride, sucrose, polysorbate 80, and water for injection.

[0102] In some embodiments, the therapeutic composition of the oncolytic virus comprises VG2025 and a pharmaceutically acceptable carrier. The phrase "pharmaceutically acceptable carrier" means any carrier, diluent, or excipient that does not interfere with the effectiveness of the oncolytic virus's biological activity and is non-toxic to the recipient (see generally Remington: 20 The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st edition (May 1, 2005) and The United States Pharmacopeia: The National Formulary (USP 40–NF 35).

[0103] In some embodiments, non-limiting examples of suitable drug carriers include phosphate-buffered saline solutions, water, emulsions (such as oil / water emulsions), various types of wetting agents, sterile solutions, etc. Other pharmaceutically acceptable carriers include gels, bioabsorbable matrix materials, implantable elements containing oncolytic viruses, or any other suitable media, delivery or dispensing tools or materials. Such carriers can be formulated using conventional methods and can be administered to the subject at an effective dose. Other pharmaceutically acceptable excipients include, but are not limited to, water, saline, polyethylene glycol, hyaluronic acid, and ethanol. Pharmaceutically acceptable salts may also be included, such as mineral salts (e.g., hydrochloride, hydrobromide, phosphate, sulfate, etc.) and salts of organic acids (e.g., acetate, propionate, malonate, benzoate, etc.). Such pharmaceutically acceptable (pharmaceutical grade) carriers, diluents, and excipients that can be used to deliver oHSV to cancer cells will preferably not induce an immune response in the individual receiving the composition (the subject) (and will preferably be administered without excessive toxicity).

[0104] The compositions can be stored at temperatures favorable for stable shelf life, including room temperature (about 20°C), 4°C, -20°C, -80°C, and in liquid nitrogen. Since compositions intended for in vivo use typically do not contain preservatives, they will generally be stored at cooler temperatures. The compositions can be stored dry (e.g., lyophilized) or in liquid form.

[0105] In this invention, the dosing regimen can be adjusted to obtain the optimal target response (e.g., treatment or prevention). For example, it can be administered as a single dose, multiple times over a period of time, or the dose can be reduced or increased proportionally according to the urgency of the treatment situation.

[0106] The typical non-limiting range of the therapeutic or preventative effective dose of the bispecific antibody of the present invention is 0.02-50 mg / kg, for example 0.1-50 mg / kg, 0.1-25 mg / kg, or 1-10 mg / kg. It should be noted that the dosage may vary depending on the type and severity of the symptoms requiring treatment. Furthermore, those skilled in the art will understand that for any given patient, a specific dosing regimen should be adjusted over time based on the patient's needs and the physician's professional evaluation; the dosage ranges given herein are for illustrative purposes only and do not limit the use or scope of the pharmaceutical compositions of the present invention.

[0107] The variable regions of the light and heavy chains determine antigen binding; each chain's variable region contains three hypervariable regions called complementarity-determining regions (CDRs). The heavy chain (H chain) CDRs include HCDR1, HCDR2, and HCDR3, while the light chain (L chain) CDRs include LCDR1, LCDR2, and LCDR3. These were named by Kabat et al., see Bethesda Md, Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication (1-3) 1991: 91-3242.

[0108] Preferably, the CDR can also be defined by the IMGT numbering system, see Ehrenmann, Francois, QuentinKaas, and Marie-Paule Lefranc. “IMGT / 3Dstructure-DB and IMGT / DomainGapAlign: a database and a tool for immunoglobulins or antibodies, T cell receptors, MHC, IgSF and MhcSF.” Nucleic acids research 38.suppl_1 (2009): D301-D307.

[0109] The VG2025 of this invention is a recombinant HSV-1 that utilizes dual regulation of transcription and translation of key viral genes to limit viral replication in tumor cells and enhance tumor-specific virulence without compromising safety. Furthermore, VG2025 expresses a payload cassette consisting of IL12, IL15, and the IL15 receptor α subunit. Payload expression is controlled by the CXCR4 promoter for tumor-specific immune stimulation. Finally, the viral glycoprotein B (gB) in VG2025 is truncated to promote viral spread in the tumor microenvironment through enhanced fusion.

[0110] In this invention, the expression of ICP34.5 in VG2025 is post-transcriptionally (translational) regulated. Briefly, in wild-type HSV-1, there are two copies of the ICP34.5 gene. In VG2025, one copy of ICP34.5 has been deleted. For the remaining ICP34.5 gene, VG2025 inserts multiple copies of the binding domains of miR-124 and miR-143 into the 3'UTR region to regulate its expression post-transcriptionally.

[0111] ICP34.5 is encoded by the HSV late-stage gene g-34.5. It is known to suppress the antiviral immunity of host cells (especially neurons), thereby causing neurotoxicity. To eliminate the function of ICP34.5 in neurons and other normal cells, while preserving its activity in tumor cells for robust replication, VG2025 uses microRNA as a post-transcriptional control, rather than gene deletion or using a specific promoter, to control ICP34.5 expression, thus achieving differential expression of ICP34.5 in tumor cells. In short, microRNA (also known as “miRNA” or “miR”) is... A 22-nucleotide non-coding small RNA encoded by a miRNA gene is transcribed by RNA polymerase II to produce a primary miRNA (primiRNA). Mature single-stranded (ss) miRNAs form a miRNA-associated RNA-induced silencing complex (miRISC). The miRNA in the miRISC can influence gene expression by binding to the 3'-untranslated region (3'-UTR) of the target mRNA. This region consists of sequences recognized by the miRNA. If the complementarity of the miRNA:mRNA complex is perfect, the mRNA is degraded by Ago2, a protein belonging to the Argonaute family. However, if the complementarity is imperfect, translation of the target mRNA is not completely degraded but rather repressed.

[0112] miRNAs are expressed differentially in tissue-specific forms. One example is miR-124. Although the precursors of miR-124 differ from species, the mature miR-124 sequences in humans, mice, and rats are identical. MiR-124 is the most abundant miRNA expressed in neuronal cells and is also highly expressed in immune cells and organs (Qin et al., 2016, miRNA-124 in immune system and immune disorders. Frontiers in Immunology, 7(OCT), 1-8). Another example of differential expression of miRNAs is miR-143 (Lagos-Quintana et al., 2002, Identification of tissue-specific MicroRNAs from mouse. Current Biology, 12(9), 735-739. MiR-143 is constitutively expressed in normal tissues but is significantly downregulated in cancer cells (Michael et al., 2003, Reduced Accumulation of Specific MicroRNAs in Colorectal Neoplasia. Molecular Cancer Research, 1(12), 882-892).

[0113] The 3'UTR region of the ICP34.5 gene in VG2025 contains multiple copies of binding domains that are fully complementary to miR-124 and miR-143 (also referred to as "miRNA target sequences," "miRNA binding sequences," or "miRNA binding sites"). Binding of miR-124 and miR-143 to the 3'UTR of ICP34.5 mRNA leads to mRNA degradation; therefore, this gene is downregulated post-transcriptionally in normal cells but not in tumor cells. This design allows for differential expression of ICP34.5 in tumor cells.

[0114] HSV-1 viral replication depends on a cascade of viral gene expression, in which immediate early gene products (especially ICP4 and ICP27) control the subsequent expression of early and late viral genes, which in turn control the viral lysis replication cycle. The absence of ICP4 or ICP27 leads to complete cessation of viral replication and a significant reduction in viral gene expression, making ICP4 and ICP27 excellent targets for tumor-specific regulation in oncolytic HSV.

[0115] While ICP4 is the primary transcription factor regulating viral gene expression, ICP27 is a multifunctional protein that regulates the transcription of many viral genes. ICP27 plays a role in all stages of mRNA biogenesis, from transcription, RNA processing and export to translation. ICP27 is also involved in nuclear protein quality control, cell cycle control, activation of stress signaling pathways, and prevention of apoptosis.

[0116] In VG2025 of this invention, the natural promoter of ICP27 is replaced by a 432bp promoter of human carcinoembryonic antigen (CEA) (Beauchemin and Arabzadeh, 2013, Carcinoembryonic antigen-related celladhesion molecules (CEACAMs) in cancer progression and metastasis. Cancer and Metastasis Reviews, 32(3–4), 643–671; Hammarström 1999, The carcinoembryonic antigen (CEA) family. Structures, suggested functions and expression in normal and malignant tissues. Seminars in cancer biology 9 (2), pp. 67–81; Kodera et al., 1993, Expression of carcinoembryonic antigen (CEA) and nonspecific crossreacting antigen (NCA) in gastrointestinal cancer; the correction with degress of differentiation. Br. J Cancer 68 (1), pp. 130–136). CEA belongs to a subgroup of 12 genes in a family of 22 genes called carcinoembryonic antigen cell adhesion molecules (CEACAM) (Beauchemin & Arabzadeh, 2013). CEA plays important roles in cellular processes, including inhibiting differentiation programs, suppressing anodic apoptosis and apoptosis, and disrupting cell polarization and tissue architecture (Beauchemin & Arabzadeh, 2013).

[0117] The VG2025 of this invention co-expresses IL12, IL15, and the IL15 receptor α subunit to further stimulate the immune regulatory response. IL12 expression promotes the polarization of antigen-exposed T cells towards the inflammatory and anti-tumor TH1 phenotype, while IL-15 activates NK cells to further increase tumor killing and antigen-presenting cell activation. In addition to IL15 expression, VG2025 also encodes IL15Rα to further enhance immune stimulation.

[0118] The molecules IL-12, IL-15, and IL-15Rα can have various configurations in the oHSV vector. For example, each molecule can be expressed individually from a separate promoter / regulatory region or co-expressed from one or two separate promoters / regulatory regions.

[0119] In some implementations, two or three of the molecules are expressed in a single transcript from a single promoter, and their coding sequences are separated by one or more self-cleaving peptides 2A. These peptides are short peptides (approximately 18-22 amino acids) and are inserted into frames between the coding sequences. During translation, the ribosome skips the synthesis of the glycyl-prolyl peptide bond at the C-terminus of the 2A peptide, resulting in a cleavage between the 2A peptide and its nearest adjacent downstream protein. Thus, they produce equimolar levels of multiple gene products from the same mRNA. This "cleavage" occurs between gly-pro residues at the C-terminus, meaning that the upstream cistron will have additional residues added to its C-terminus, while the downstream cistron begins with proline.

[0120] Transcription of IL-12, IL-15, and IL-15Rα is preferably driven by a single promoter (CXCR4), and the peptide is linked to a 2A self-cleaving peptide (Z. Liu et al., 2017, Systematic comparison of 2A peptides for cloning multi-genes in polycistronic vector. Scientific Reports, 7(2), 1-9), which produces three separate proteins via ribosome jumping during translation. Although the payload is intended to be expressed within the tumor, unintended expression may occur in normal tissues in cases of viral “leakage” to extratumor regions or during systemic viral delivery. To mitigate the potential risk of extratumor expression of IL12 / IL15, the expression cassette of the payload in VG2025 is driven by a single promoter of CXC chemokine receptor 4 (CXCR4) (Moriuchi et al., 1997, Cloning and analysis of the promoter region of CXCR4, a coreceptor for HIV-1 entry. Journal of Immunology (Baltimore, Md.: 1950), 159(9), 4322-429; Caruz et al., 1998, Genomic organization and promoter characterization of human CXCR4 gene. FEBS Letters, 426(2), 271-278. CXCR4 is a seven-transmembrane G protein-coupled receptor that was initially isolated from human blood mononuclear cells as a cofactor for HIV fusion and T cell entry (Moriuchi et al., 1997).

[0121] Other promoters can be used. Cellular promoters, viral promoters, etc., are suitable. The promoter can be constitutive, inducible, or cell / tissue specific. Many promoters are well-known. One specific promoter that can be used is the constitutive EF-1α promoter.

[0122] The sequence is assembled in one or more expression cassettes. Examples provide specific versions of some exemplary expression cassettes. The expression cassette can be inserted into the HSV genome at any location without disrupting critical functions (e.g., replication). In some implementations, the expression cassette is inserted into an internal or terminal repeat region after the initial deletion of the repeat region. Other suitable insertion regions include between viral genes, such as between the UL3 and UL4 viral genes, between the UL50 and UL51 genes, and between US1 and US2.

[0123] Representative examples of the selected expression cassettes / carriers are also described in CN202010720662.1, PCT Publications WO2018 / 026872 and WO2022 / 204434, which are incorporated herein by reference in their entirety.

[0124] HSV-1 membrane fusion is a critical step in infection. It relies on four essential viral glycoproteins (gB, gD, gH, and gL) that mediate entry into the host cell by fusing the viral envelope with the host cell membrane. The core fusion protein is glycoprotein B (gB), a 904-residue glycosylated transmembrane protein encoded by the UL27 gene of HSV-1. Various mutation types within the cytoplasmic domain of gB have resulted in a high fusion phenotype, increasing cell-cell fusion (Chowdary & Heldwein, 2010, Synctial Phenotype of C-Terminally Truncated Herpes SimplexVirus Type 1 gB is Associated with Diminished Membrane Interactions. Journal of Virology, 84(10), 4923-4935). In one embodiment, gB can be modified by truncating the C-terminal amino acid positions 877-904 from the full-length protein.

[0125] The present invention also includes one or more aspects selected from the following: 1. A drug combination containing an effective amount of anti-PD-1-anti-VEGFA bispecific antibody and oncolytic virus, wherein: The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; Wherein, the first protein functional region is an immunoglobulin and the second protein functional region is a single-chain antibody; or, the first protein functional region is a single-chain antibody and the second protein functional region is an immunoglobulin. in, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; or, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; And, the oncolytic virus is herpes simplex virus type 1 (HSV-1).

[0126] 2. The drug combination according to aspect 1, wherein, The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 14; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. or, The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 16; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 14.

[0127] 3. The drug combination according to any one of aspects 1 to 2, wherein: (1) The immunoglobulin is human IgG1 subtype; (2) The immunoglobulin is human IgG1 subtype, and according to the EU numbering system, the heavy chain constant region of the immunoglobulin has one of the following combinations of mutations: L234A and L235A; L234A and G237A; L235A and G237A; or L234A, L235A and G237A; (3) The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18; (4) The first protein functional region is directly connected to the second protein functional region or is connected through a first linker; and / or the heavy chain variable region of the single-chain antibody is directly connected to the light chain variable region of the single-chain antibody or is connected through a second linker; (5) The first protein functional region is directly connected to the second protein functional region or is connected through a first linker; and / or the heavy chain variable region of the single chain antibody is directly connected to the light chain variable region of the single chain antibody or is connected through a second linker, and the first linker and the second linker are independently polypeptides of n peptides shown in SEQ ID NO: 19 in series, where n is 1, 2, 3, 4, 5 or 6; (6) The first protein functional region is directly connected to the second protein functional region or connected through a first linker; and / or the heavy chain variable region of the single-chain antibody is directly connected to the light chain variable region of the single-chain antibody or connected through a second linker; and the first linker and the second linker are independently polypeptides as shown in SEQ ID NO: 20; (7) The single-chain antibody is independently connected in sequence as follows: heavy chain variable region-linker-light chain variable region (VH-linker-VL), or light chain variable region-linker-heavy chain variable region (VL-linker-VH). (8) The amino acid sequence of the single-chain antibody is shown in SEQ ID NO: 21; (9) The first protein functional region and the second protein functional region are independently one, two or more; (10) The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; (11) The bispecific antibody is a bispecific antibody in the form of IgG-scFv; and / or (12) The bispecific antibody is a tetravalent bispecific antibody in the form of IgG-scFv.

[0128] 4. The drug combination according to any one of aspects 1 to 3, wherein the anti-PD-1-anti-VEGFA bispecific antibody is selected from the following 1)-7). 1) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; 2) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; 3) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; 4) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; 5) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; 6) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains are independently connected by one or more disulfide bonds; 7) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains.

[0129] 5. The drug combination according to any one of aspects 1 to 4, wherein the oncolytic virus is selected from: (1) Modified HSV-1; (2) Oncolytic virus VG2025; (3) Modified wild-type HSV-1; (4) Modified wild-type HSV-1 strain 17+; (5) Oncolytic viruses, whose ICP34.5 levels are lower in normal cells than in tumor cells; (6) Oncolytic viruses, whose ICP34.5 levels are lower in normal nerve cells than in tumor cells; (7) Modified HSV-1, and compared with wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the second copy is downregulated at the posttranscriptional mRNA level; (8) Modified HSV-1, in which the first copy of the two copies of the ICP34.5 gene is deleted or inactivated relative to wild-type HSV-1, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences. (9) Modified HSV-1, wherein, relative to wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences; the miRNA target sequences target miR-124 and / or miR-143; (10) Modified HSV-1, wherein, relative to wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences; said miRNA target sequences target miR-124 and / or miR-143; the sequence of miR-124 is shown in SEQ ID NO: 30; the sequence of miR-143 is shown in SEQ ID NO: 31; (11) Modified HSV-1, and the natural promoter of ICP27 was replaced by a tumor-specific promoter; (12) Modified HSV-1, and the natural promoter of ICP27 is replaced by the human carcinoembryonic antigen promoter; (13) Modified HSV-1, and the natural promoter of ICP27 is replaced by the human carcinoembryonic antigen promoter; the sequence of the human carcinoembryonic antigen promoter is shown in SEQ ID NO: 23; (14) A modified HSV-1, and comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; (15) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. (16) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; the expression cassette being driven by a CXCR4 promoter; (17) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (18) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; the expression cassette being driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25; (19) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25. (20) Modified HSV-1, and comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide. (21) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (22) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; (23) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. (24) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (25) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (26) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25; (27) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25. (28) Modified HSV-1, and the C-terminal amino acid positions 877-904 of glycoprotein B were truncated; (29) Modified HSV-1, and the C-terminal amino acid positions 877-904 of glycoprotein B were truncated; the amino acid sequence of the truncated glycoprotein B is shown in SEQ ID NO: 24; (30) Modified wild-type HSV-1 or modified wild-type HSV-1 strain 17+, and possessing any one, two, three, or all four of the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (31) Any one or any combination of the above items.

[0130] 6. The drug combination according to any one of aspects 1 to 5 further comprises an effective amount of a chemotherapy drug, wherein the chemotherapy drug is a chemotherapy drug of the mFOLFOX6 regimen, the FOLFIRI regimen, or the CapeOX regimen.

[0131] 7. The drug combination according to any one of aspects 1 to 6, comprising an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody and an oncolytic virus, or comprising an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus and a chemotherapeutic agent, and selected from any one of (1) to (15) below: (1) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (2) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (3) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (4) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (5) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (6) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (7) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (8) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (10) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (11) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; (12) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO: 14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; (13) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; (14) The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first functional region of VEGFA protein, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO: 16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; (15) The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO: 26. The amino acid sequence of IL15 is shown in SEQ ID NO: 27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine.

[0132] 8. The drug combination according to any one of aspects 1 to 7, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody is mixed with an oncolytic virus, and the drug combination is a pharmaceutical composition; or The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are packaged in separate containers, and the drug combination is a combination product. (2) Based on the mass of the anti-PD-1-anti-VEGFA bispecific antibody, the unit dose of the anti-PD-1-anti-VEGFA bispecific antibody is 100mg-2500mg, 100mg-2000mg, 100mg-1500mg, 100mg-1200mg, 100mg-1000mg, 200mg-800mg, 200mg-500mg, 300mg-600mg, 400mg-500mg, or 450mg; (3) The titer of the oncolytic virus is approximately 5 × 10⁻⁶. 4 pfu / mL - approximately 2 × 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 107 pfu / mL, approximately 10 7 pfu / mL - approximately 10 8 pfu / mL, approximately 10 8 pfu / mL - approximately 10 9 pfu / mL or approximately 10 8 pfu / mL; (4) The drug combination further comprises one or more pharmaceutically acceptable excipients; (5) The drug combination also includes a product instruction manual.

[0133] 9. The combination of drugs according to any one of aspects 1 to 8, used for the treatment or prevention of tumors.

[0134] 10. The drug combination according to aspect 9, wherein the tumor is selected from one or more of the following: Colorectal cancer, liver cancer, advanced tumors, metastatic tumors, colorectal cancer liver metastases, advanced colorectal cancer liver metastases, hepatocellular carcinoma, advanced liver cancer, and advanced hepatocellular carcinoma.

[0135] 11. The drug combination according to any one of aspects 1 to 10, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Alternatively, the anti-PD-1-anti-VEGFA bispecific antibody may be administered after the oncolytic virus, with an interval of 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg or 1200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, or administration once on day 1 and day 3 respectively, and no administration is given during the rest of the cycle; and each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or oral administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) A combination of any of the items selected from (1)-(8) above; (10) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. (11) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion; (12) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. or (13) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, by intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion.

[0136] 12. Use of the pharmaceutical combination described in any one of aspects 1 to 11 in the preparation of a medicament for treating or preventing tumors.

[0137] 13. The use according to aspect 12, wherein the tumor is selected from one or more of the following: Colorectal cancer, liver cancer, advanced tumors, metastatic tumors, colorectal cancer liver metastases, advanced colorectal cancer liver metastases, hepatocellular carcinoma, advanced liver cancer, and advanced hepatocellular carcinoma.

[0138] 14. The use according to any one of aspects 12 to 13, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Alternatively, the anti-PD-1-anti-VEGFA bispecific antibody may be administered after the oncolytic virus, with an interval of 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg or 1200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, or administration once on the first and third days of each cycle, and no administration is given during the rest of the cycle; and each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or oral administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) A combination of any of the items selected from (1)-(8) above; (10) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. (11) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion; (12) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. or (13) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, by intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion.

[0139] 15. A method of treating or preventing tumors, comprising the step of administering to a subject in need an effective amount of the combination of drugs described in any one of aspects 1 to 11.

[0140] 16. The method according to aspect 15, wherein the tumor is selected from one or more of the following: Colorectal cancer, liver cancer, advanced tumors, metastatic tumors, colorectal cancer liver metastases, advanced colorectal cancer liver metastases, hepatocellular carcinoma, advanced liver cancer, and advanced hepatocellular carcinoma.

[0141] 17. The method according to any one of aspects 15 to 16, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Alternatively, the anti-PD-1-anti-VEGFA bispecific antibody may be administered after the oncolytic virus, with an interval of 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg or 1200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 8 days, 10 days, 15 days, 22 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, or administration once on the first and third days of each cycle, and no administration is given during the rest of the cycle; and each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or oral administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) A combination of any of the items selected from (1)-(8) above; (10) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. (11) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion; (12) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. or (13) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, by intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion.

[0142] In this invention, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the cell culture, molecular genetics, nucleic acid chemistry, and immunology laboratory procedures used herein are all standard procedures widely used in their respective fields. To better understand this invention, definitions and explanations of relevant terms are provided below.

[0143] As used herein, when referring to the amino acid sequence of a VEGFA protein, it includes the full-length VEGFA protein, as well as fusion proteins of VEGFA, such as fragments fused to the Fc protein fragments (mFc or hFc) of mouse or human IgG. However, those skilled in the art will understand that mutations or variations (including, but not limited to, substitutions, deletions, and / or additions) can be naturally generated or artificially introduced into the amino acid sequence of a VEGFA protein without affecting its biological function. Therefore, in this invention, the term "VEGFA protein" should include all such sequences, including their natural or artificial variants. Furthermore, when describing a sequence fragment of a VEGFA protein, it also includes the corresponding sequence fragment from its natural or artificial variants.

[0144] As used herein, when referring to the amino acid sequence of the PD-1 protein, it includes the full-length PD-1 protein, or the extracellular fragment PD-1ECD of PD-1, or a fragment containing PD-1ECD; it also includes fusion proteins of PD-1ECD, such as fragments fused with a fragment of the Fc protein (mFc or hFc) of mouse or human IgG. However, those skilled in the art will understand that mutations or variations (including, but not limited to, substitutions, deletions, and / or additions) can be naturally generated or artificially introduced into the amino acid sequence of the PD-1 protein without affecting its biological function. Therefore, in this invention, the term "PD-1 protein" should include all such sequences, including its natural or artificial variants. Furthermore, when describing a sequence fragment of the PD-1 protein, it also includes the corresponding sequence fragment from its natural or artificial variants.

[0145] As used in this article, the term EC 50 The half-maximal effect concentration refers to the concentration that produces a 50% maximum effect.

[0146] As used herein, the term "antibody" refers to an immunoglobulin molecule typically composed of two pairs of polypeptide chains (each pair consisting of one "light" (L) chain and one "heavy" (H) chain). In a general sense, the heavy chain can be understood as the larger polypeptide chain in an antibody, and the light chain as the smaller polypeptide chain. Light chains can be classified as κ and λ light chains. Heavy chains are typically classified as μ, δ, γ, α, or ε, and antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within both light and heavy chains, variable and constant regions are linked by a "J" region of approximately 12 or more amino acids, and the heavy chain also contains a "D" region of approximately 3 or more amino acids. Each heavy chain is further divided by a heavy chain variable region (V... H ) and heavy chain constant region (C H The heavy chain constant region consists of three structural domains (C). H1 C H2 and C H3 It consists of ) light chains. Each light chain is composed of a light chain variable region (V L ) and light chain constant region (C L It consists of a light chain constant region composed of a structural domain C. L Composition. The constant region of an antibody mediates the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. V H and V L The region can be further subdivided into highly variable regions (called complementary determinant regions (CDRs)), interspersed with more conservative regions called framework regions (FRs). Each V H and VL It consists of three CDRs and four FRs arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4, from the amino terminus to the carboxyl terminus. The variable region (V) of each heavy chain / light chain pair... H and V L The amino acids are distributed to the respective regions or domains to form antibody binding sites. The allocation of amino acids to each region or domain follows the definitions in Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883. In particular, the heavy chain may also contain more than three CDRs, such as six, nine, or twelve. For example, in the bispecific antibody of this invention, the heavy chain may be the C-terminus of the heavy chain of an IgG antibody linked to the scFv of another antibody, in which case the heavy chain contains nine CDRs. The term "antibody" is not limited to any particular method of antibody production. For example, it includes, in particular, recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. Antibodies can be different isotypes of antibodies, such as IgG (e.g., IgG1, IgG2, IgG3 or IgG4 subtypes), IgA1, IgA2, IgD, IgE or IgM antibodies.

[0147] Antigen-binding fragments of an antibody (e.g., the antibody fragments described above) can be obtained from a given antibody using conventional techniques known to those skilled in the art (e.g., recombinant DNA techniques or enzymatic or chemical fragmentation methods), and the antigen-binding fragments of the antibody can be screened for specificity in the same manner as those used for intact antibodies.

[0148] In this article, unless the context clearly indicates otherwise, when referring to the term "antibody," it includes not only the complete antibody but also the antigen-binding fragment of the antibody.

[0149] As used herein, the terms “monoclonal antibody” and “monoclonal antibody” refer to an antibody or a fragment of an antibody derived from a group of highly homologous antibody molecules—that is, a group of identical antibody molecules except for the possibility of spontaneous natural mutations. Monoclonal antibodies exhibit high specificity for a single epitope on an antigen. Polyclonal antibodies, as opposed to monoclonal antibodies, typically contain at least two or more different antibodies that typically recognize different epitopes on an antigen. Monoclonal antibodies are usually obtained using hybridoma techniques first reported by Kohler et al. (Kohler et al., Nature, 256:495, 1975), but can also be obtained using recombinant DNA techniques (see US Patent 4,816,567).

[0150] The term "oncolytic herpesvirus" or "oHSV" usually refers to a virus that can replicate in tumor cells.

[0151] Herpes simplex virus that replicates and kills tumor cells. The term "oncolytic herpes simplex virus" or "oHSV" refers to a herpes simplex virus that can replicate and kill tumor cells.

[0152] As used herein, the terms “pharmaceuticalally acceptable excipient” or “pharmaceuticalally acceptable carrier and / or excipient” mean a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and the active ingredient, which is well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by GennaroAR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and include, but are not limited to: pH adjusters, surfactants, adjuvants, and ionic strength enhancers. For example, pH adjusters include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic, or nonionic surfactants, such as Tween-80; and ionic strength enhancers include, but are not limited to, sodium chloride.

[0153] As used herein, the term "effective amount" means an amount sufficient to achieve, or at least partially achieve, the desired effect. For example, an effective amount for disease prevention is an amount sufficient to prevent, stop, or delay the onset of disease; an effective amount for disease treatment is an amount sufficient to cure or at least partially stop the disease and its complications in a patient already suffering from the disease. Determining such an effective amount is entirely within the capabilities of those skilled in the art. For example, an effective amount for therapeutic purposes will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general characteristics such as age, weight, and sex, the manner of administration of the drug, and other concurrent treatments, etc.

[0154] As used herein, the term "unit dose" refers to the dose of active ingredient contained in the smallest unit of packaging containing a certain amount of medicine, for example, the dose of antibody contained in a vial of antibody injection is a unit dose.

[0155] As used herein, the terms “subject,” “patient,” or “subject” are used interchangeably. “Subject,” “patient,” or “subject” includes any human or non-human animal. The term “non-human animal” includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In some embodiments, the terms “subject,” “patient,” or “subject” are mammals. In some embodiments, the subject, patient, or subject is a mouse. In some embodiments, the subject, patient, or subject is a human.

[0156] As used herein, “combined use” or “concurrent use” means that two or more active substances may be administered to a subject simultaneously, or sequentially, as single formulations, in any order. Alternatively, all of the active ingredients may be formulated in a single formulation and administered to a subject simultaneously. Alternatively, portions of the active ingredients may be formulated in a single formulation, and other portions of the active ingredients may be administered to a subject simultaneously, or sequentially, in any order, as single formulations.

[0157] "Recurrent" cancer is cancer that recurs at the initial site or a distant site after a response to initial treatment (such as surgery). "Locally recurrent" cancer is cancer that recurs at the same location as the previously treated cancer after treatment.

[0158] Metastatic cancer refers to cancer that has spread from one part of the body (such as the lungs) to another part of the body.

[0159] As used in this article, the term "RECIST 1.1 efficacy criteria" refers to the definition of target or non-target damage based on the context of the measured response, as stated by Eisenhauver et al., E.A. et al., Eur. J Cancer 45:228-247 (2009). Prior to immunotherapy, it was the most commonly used standard for evaluating the efficacy of treatment in solid tumors. Specifically, it includes the following aspects: 1. Selection and measurement of target lesions: The RECIST 1.1 criteria require the selection of up to five target lesions for measurement, with a maximum of two target lesions per organ. The longest diameter of the target lesion must be greater than or equal to 10 mm (15 mm or greater for lymph nodes). The longest diameter of the target lesion is measured using imaging examinations (such as CT, MRI), and the baseline value is recorded. 2. Assessment of non-target lesions: Non-target lesions refer to other lesions that were not selected as target lesions. The RECIST 1.1 criteria require a qualitative assessment of non-target lesions, recording whether they have progressed, stabilized, or regressed. 3. Efficacy Assessment: Based on changes in target and non-target lesions, the RECIST 1.1 criteria categorize efficacy into four types: complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD). 4. Follow-up and Re-examination: The RECIST 1.1 criteria require regular imaging examinations, typically every 6-8 weeks, to assess tumor changes. Treatment plans are adjusted based on follow-up results. 5. Management of Special Cases: For certain special cases, such as intratumoral hemorrhage or necrosis, the RECIST 1.1 criteria also provide corresponding management methods. A comprehensive assessment of tumor changes is necessary, taking into account the actual clinical situation. In 2016, the RECIST working group revised the existing "RECIST v.1.1" and proposed a new set of criteria, namely the "irRECIST criteria" described in this article, aiming to better evaluate the efficacy of immunotherapy drugs.

[0160] As used in this article, the term "ECOG" scoring system is an indicator of a patient's general health status and tolerance to treatment based on their physical performance. The ECOG performance status scoring system uses a scale of 0, 1, 2, 3, 4, and 5. A score of 0 indicates completely normal activity levels, with no difference from pre-illness activity levels. A score of 1 indicates the ability to walk freely and perform light physical activities, including general housework or office work, but not strenuous physical activity.

[0161] As used in this article, the terms “complete remission (CR)”, “partial remission (PR)”, “stable disease (SD)”, “progressive disease (PD)”, “objective response rate (ORR)”, and “disease control rate (DCR)” are defined as follows: When the disease is a solid tumor, the efficacy of treatment for solid tumors is evaluated according to the following criteria (see New response evaluation criteria in solid tumors: Revised RECIST guideline (version 1.1), EAEisenhauer et al., *EUROPEAN JOURNAL OF CANCER*, 45 (2009), pp. 228-247): "Complete Response (CR)," "Partial Response (PR)," "Stable Disease (SD)," "Progressive Disease (PD)," "Objective Response Rate (ORR)," and "Disease Control Rate (DCR)." The specific evaluation of target lesions is as follows: Complete remission (CR): All target lesions disappear, the short diameter of all pathological lymph nodes (including target nodules and non-target nodules) must be reduced to <10mm, and there are no new lesions.

[0162] Partial remission (PR): The sum of the diameters of target lesions (shortest diameter of lymph nodes) is reduced by at least 30% from baseline. There is no significant progression of non-target lesions and no new lesions.

[0163] Disease stability (SD): The degree of reduction in target lesions does not reach the PR level, nor does the degree of increase reach the PD level; it falls between the two. The minimum value of the sum of diameters can be used as a reference in studies.

[0164] Disease progression (PD): The minimum sum of the diameters of all measured target lesions throughout the entire trial study is used as a reference, with a relative increase of at least 20% in the sum of diameters (or the baseline value if the baseline measurement is the minimum); in addition, the absolute value of the sum of the measured target lesion diameters must increase by at least 5 mm (the appearance of one or more new lesions is also considered disease progression).

[0165] Objective response rate (ORR): This refers to the proportion of patients whose tumors shrink to a certain extent, including cases of complete remission (CR) and partial response (PR). Subjects must have measurable tumor lesions at baseline, and the efficacy assessment criteria are based on RECIST 1.1.

[0166] Disease control rate (DCR): The percentage of patients who achieved complete remission (CR), partial remission (PR), or stable disease (SD) out of the total number of patients analyzed.

[0167] PFS and OS can be measured according to the standards set by the National Cancer Institute and the U.S. Food and Drug Administration for new drug approval. See Johnson et al., J. Clin. Oncol. 21(7):1404-1411 (2003).

[0168] Progression-free survival (PFS) refers to the time from enrollment to disease progression or death. PFS is generally measured using the Kaplan-Meier method and the Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 criteria. In general, progression-free survival refers to the state in which a patient remains alive and whose cancer has not progressed.

[0169] Overall survival (OS) refers to the time from patient enrollment to death or when the last known date of survival is determined. OS includes the extension of life expectancy compared to untreated or untreated individuals or patients. Overall survival refers to the state in which a patient remains alive for a specified period of time, such as one year, five years, etc., from the time of diagnosis or treatment. In a patient population, overall survival is measured as median overall survival (mOS).

[0170] Depth of Response (DpR): Depth of response refers to the degree of reduction in tumor volume or disease progression during treatment. Specifically, it is the percentage of the maximum tumor shrinkage achieved during treatment compared to baseline. If complete remission (CR) occurs, the DpR is 100. It is usually measured based on imaging or clinical assessment.

[0171] Duration of Response (DoR): Duration of response refers to the duration of tumor remission after treatment, specifically the time from the first confirmed complete response (CR) or confirmed partial response (PR) to the first disease progression for subjects who have achieved a confirmed complete response (CR) or confirmed partial response (PR).

[0172] Time to Response (TTR): The time to response refers to the time from the start of treatment to when the patient first achieves a state of remission (CR or PR).

[0173] In this invention, unless otherwise specified, the terms "first" (e.g., first protein functional region, first linker, first copy, or first peptide chain) and "second" (e.g., second protein functional region, second linker, second copy, or second peptide chain) are used for distinguishing references or clarifying expressions, and do not have a typical sequential meaning.

[0174] In this invention, unless otherwise specified, “about” or “approximately” means fluctuating within 10%, 20% or 30% of the modified numerical value or physical quantity. For example, about 100 minutes or approximately 100 minutes can be 90-110 minutes, 80-120 minutes or 70-130 minutes.

[0175] Beneficial effects of the invention

[0176] The present invention achieves any one or more of the technical effects described in items (1) to (4) below: (1) The drug combination of the present invention can effectively prevent or treat tumors.

[0177] (2) The combination of anti-PD-1-anti-VEGFA bispecific antibody Ivonescimab and oncolytic virus VG2025 has a pharmacological effect of effectively promoting the expression of cytokines such as IL-2 / IFN-γ / TNF-α and inhibiting tumor growth. The effect is better than that of anti-PD-1-anti-VEGFA bispecific antibody Ivonescimab or oncolytic virus VG2025 alone, achieving a synergistic effect.

[0178] (3) The drug combination of the present invention can improve the survival of the treated patients (e.g., median survival, progression-free survival or overall survival) compared with any single drug administered alone.

[0179] (4) The drug combination of the present invention has better efficacy in at least one aspect of objective response rate, disease control rate, tolerability and side effects compared with any one of the drugs administered alone. Attached Figure Description

[0180] Figure 1A Anti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote IL-2 secretion by PBMCs (LS174T cell system).

[0181] Figure 1B Anti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote IFN-γ secretion by PBMCs (LS174T cell system).

[0182] Figure 1C Anti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote TNF-α secretion by PBMCs (LS174T cell system).

[0183] Figure 2AAnti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote IL-2 secretion by PBMCs (LS1034 cell system).

[0184] Figure 2B Anti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote IFN-γ secretion by PBMCs (LS1034 cell system).

[0185] Figure 2C Anti-PD-1 and anti-VEGFA bispecific antibodies combined with oncolytic virus VG2025 promote TNF-α secretion by PBMCs (LS1034 cell system).

[0186] The sequences involved in this invention are shown in Table A below.

[0187] Table A: Partial Sequences Involved in the Invention Detailed Implementation

[0188] The embodiments of the present invention will be described in detail below with reference to examples. Those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the examples, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased on the market.

[0189] Preparation Example 1: Preparation of bispecific antibodies against PD-1 and anti-VEGFA or their antigen-binding fragments

[0190] The preparation of the antibody Ivonescimab is described in US Patent Publication US2023027029 A1 (PCT Publication WO2021104302 A1), which is hereby incorporated herein by reference. Ivonescimab is a bispecific antibody against PD-1 and against VEGFA, comprising a first protein functional region targeting VEGFA and a second protein functional region targeting PD-1. By constructing a bispecific antibody with a humanized IgG1-ScFv structure, a bispecific antibody with a tetravalent structure is generated, binding VEGF-A at the immunoglobulin end and PD-1 at the single-chain antibody end, enabling it to simultaneously block PD-1 and VEGF-A signaling. The relevant sequences are as follows: The amino acid sequence of the heavy chain variable region of the immunoglobulin moiety is shown in SEQ ID NO: 13. The amino acid sequence of the light chain variable region of the immunoglobulin moiety is shown in SEQ ID NO: 14.

[0191] The amino acid sequence of the heavy chain variable region of the single-chain antibody moiety is shown in SEQ ID NO: 15. The amino acid sequence of the light chain variable region of the single-chain antibody moiety is shown in SEQ ID NO: 16.

[0192] According to the EU numbering system, a point mutation from leucine to alanine (L234A) was introduced at position 234 in its heavy chain hinge region, and a point mutation from leucine to alanine (L235A) was introduced at position 235.

[0193] The amino acid sequence of the heavy chain of the immunoglobulin moiety is shown in SEQ ID NO: 17. The amino acid sequence of the light chain of the immunoglobulin moiety is shown in SEQ ID NO: 18.

[0194] The amino acid sequence of the linker repeat unit is shown in SEQ ID NO: 19. The amino acid sequence of the linker is shown in SEQ ID NO: 20.

[0195] The amino acid sequence of the single-chain antibody is shown in SEQ ID NO: 21.

[0196] The amino acid sequence of the heavy chain of Ivonescimab is shown in SEQ ID NO: 22. The amino acid sequence of the light chain of Ivonescimab is shown in SEQ ID NO: 18.

[0197] Using techniques well-known to those skilled in the art, such as analyzing the amino acid sequence of the CDR region of the bispecific antibody of the present invention according to the IMGT definition using the VBASE2 database, the results are as follows: (1) Anti-PD-1-anti-VEGFA bispecific antibody (Evoxi) immunoglobulin fraction The amino acid sequences of the three CDR regions of its heavy chain variable region are as follows: HCDR1: GYTFTNYG (SEQ ID NO: 1) HCDR2: INTYTGEP (SEQ ID NO: 2) HCDR3: AKYPHYYGSSHWYFDV (SEQ ID NO: 3) The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1: QDISNY (SEQ ID NO: 4) LCDR2: FTS (SEQ ID NO: 5) LCDR3: QQYSTVPWT (SEQ ID NO: 6) (2) Anti-PD-1-anti-VEGFA bispecific antibody (Evoxi) single-chain antibody portion The amino acid sequences of the three CDR regions of its heavy chain variable region are as follows: HCDR1: GFAFSSYD (SEQ ID NO: 7) HCDR2: ISGGGRYT (SEQ ID NO: 8) HCDR3: ANRYGEAWFAY (SEQ ID NO: 9) The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1: QDINTY (SEQ ID NO: 10) LCDR2: RAN (SEQ ID NO: 11) LCDR3: LQYDEFPLT (SEQ ID NO: 12) (3) Anti-PD-1-anti-VEGFA bispecific antibody (Evoxi) The amino acid sequences of the nine CDR regions of its heavy chain are as follows: HCDR1: GYTFTNYG (SEQ ID NO: 1) HCDR2: INTYTGEP (SEQ ID NO: 2) HCDR3: AKYPHYYGSSHWYFDV (SEQ ID NO: 3) HCDR4: GFAFSSYD (SEQ ID NO: 7) HCDR5: ISGGGRYT (SEQ ID NO: 8) HCDR6: ANRYGEAWFAY (SEQ ID NO: 9) HCDR7: QDINTY (SEQ ID NO: 10) HCDR8: RAN (SEQ ID NO: 11) HCDR9: LQYDEFPLT (SEQ ID NO: 12) The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1: QDISNY (SEQ ID NO: 4) LCDR2: FTS (SEQ ID NO: 5) LCDR3: QQYSTVPWT (SEQ ID NO: 6).

[0198] Preparation Example 2: Design and Preparation of Oncolytic Virus VG2025

[0199] The preparation of the oncolytic virus VG2025 was described in WO2022 / 204434, which is hereby incorporated herein by reference. VG2025 is a recombinant HSV-1 that utilizes dual regulation of transcription and translation of key viral genes to restrict viral replication to tumor cells and enhance tumor-specific virulence without compromising safety. Furthermore, VG2025 expresses a payload cassette consisting of IL12, IL15, and the IL15 receptor α subunit. Payload expression is controlled by the CXCR4 promoter for tumor-specific immune stimulation. Finally, the viral glycoprotein B (gB) in VG2025 is truncated to promote viral spread in the tumor microenvironment through enhanced fusion. The expression control mechanisms in VG2025 are designed to increase safety without sacrificing efficacy. Specific modifications to wild-type HSV-1 strain 17+ are shown in Table 1 below.

[0200]

[0201] CEA = Carcinoembryonic antigen; CXCR4 = CXC motif chemokine receptor 4; gB = Glycoprotein B; HSV-1 = Herpes simplex virus-1; ICP27 = Infected cell polypeptide 27; ICP34.5 = Infected cell polypeptide 34.5; IL = Interleukin; miR = MicroRNA; Rα = Receptor α; TRL = Long terminal repeat; IRL = Long internal repeat; TRS = Short terminal repeat; IRS = Short internal repeat; UL = Unique long repeat.

[0202] 1. Post-transcriptional (translation) regulation

[0203] In VG2025, ICP34.5 expression is post-transcriptionally (translational) regulated. In short, wild-type HSV-1 contains two copies of the ICP34.5 gene. In VG2025, one copy of ICP34.5 has been deleted. For the remaining ICP34.5 gene, VG2025 inserts multiple copies of the binding domains of miR-124 and miR-143 into the 3'UTR region to regulate its expression post-transcriptionally.

[0204] An example of the miR-124 sequence is shown in SEQ ID NO: 30.

[0205] An example of the miR-143 sequence is shown in SEQ ID NO: 31.

[0206] The 3'UTR region of the ICP34.5 gene in VG2025 contains multiple copies (also referred to as "miRNA target sequence," "miRNA binding sequence," or "miRNA binding site"; preferably 5 copies) of a binding domain that is fully complementary to miR-124 and miR-143. Binding of miR-124 and miR-143 to the 3'UTR of ICP34.5 mRNA leads to mRNA degradation; therefore, this gene is downregulated post-transcriptionally in normal cells but not in tumor cells. This design allows for differential expression of ICP34.5 in tumor cells.

[0207] 2. ICP27 expression in VG2025 is transcriptionally controlled.

[0208] In VG2025, the natural promoter of ICP27 is replaced by a 432bp promoter of human carcinoembryonic antigen (CEA), the nucleic acid sequence of which is shown in SEQ ID NO: 23.

[0209] 3. The effective payload expression of VG2025 is tumor-enhanced.

[0210] VG2025 co-expresses IL12, IL15, and the IL15 receptor α subunit to further stimulate the immune regulatory response. IL12 expression promotes the polarization of antigen-exposed T cells towards the inflammatory and anti-tumor TH1 phenotype, while IL-15 activates NK cells to further increase tumor killing and antigen-presenting cell activation. In addition to IL15 expression, VG2025 also encodes IL15Rα to further enhance immune stimulation. For example, human IL12 may contain the amino acid sequence shown in SEQ ID NO: 26; human IL15 may contain the amino acid sequence shown in SEQ ID NO: 27; and the human IL15 receptor α subunit may contain the amino acid sequence shown in SEQ ID NO: 29.

[0211] Transcription of IL-12, IL-15 and IL-15Rα is driven by a single promoter (CXCR4), and in the polypeptide obtained by tandemly IL-12, IL-15 and IL-15Rα, the N-terminus of IL-15 and IL-15Rα are linked to 2A self-cleaving peptides, which may contain the amino acid sequence shown in SEQ ID NO: 28.

[0212] The expression cassette of the payload in VG2025 is driven by a single promoter of CXC chemokine receptor 4 (CXCR4), the nucleic acid sequence of which is shown in SEQ ID NO: 25.

[0213] Representative examples of the selected expression cassettes / carriers are also described in PCT Publications WO2018 / 026872 and WO2022 / 204434, which are incorporated herein by reference in their entirety.

[0214] 4. Truncated glycoprotein B (GB)

[0215] The core fusion protein is glycoprotein B (gB), a 904-residue glycosylated transmembrane protein encoded by the UL27 gene of HSV-1. In VG2025, gB can be modified by truncating the C-terminal amino acids from positions 877 to 904 of the full-length protein. For example, the amino acid sequence of the truncated glycoprotein B is shown in SEQ ID NO: 24.

[0216] Experimental Example 1: Combination of anti-PD-1 and anti-VEGFA bispecific antibody with oncolytic virus VG2025 promotes cytokine activity. secretion

[0217] This study investigated the pharmacological activity of the anti-PD-1-anti-VEGFA bispecific antibody Ivonescimab in combination with oncolytic virus VG2025 in promoting the secretion of cytokines IL-2, INF-γ, and TNF-α.

[0218] PBMC pretreatment: Two days before plating, routinely resuscitated PBMCs (donated by healthy volunteers) were cultured in complete medium (RPMI 1640 + 10% FBS) at 37°C in a 5% CO2 incubator; after 2 hours, once the PBMCs had recovered, SEB (Toxin Technology, catalog number: BT202) with a final concentration of 0.5 μg / mL was added for two days to activate them.

[0219] Oncolytic virus treatment of tumor cells: Two days before plating, human colon cancer cells were treated with LS174T (Shanghai Yaji Biotechnology Co., Ltd., catalog number: YS883C) and LS1034 (Nanjing Kebai Biotechnology Co., Ltd., catalog number: CBP60013) at a concentration of 3.6 × 10⁻⁶. 4 Cells / well were seeded into 96-well plates with a flat bottom and incubated overnight at 37°C. The next day, 50 μL of prepared VG2025 virus solution (Shanghai Funogen Biotechnology Co., Ltd., batch number: C032-20210302) (MOI of 1) was added to the culture wells inoculated with tumor cells and incubated at 37°C for 24 hours.

[0220] On the day of plate formation, the supernatant from virus incubation was removed, and the cells were washed twice with RPMI 1640 basal medium. PBMC (effect-to-target ratio of 5:1), Ivonescimab (Kangfang Bio, batch number: WS112-002) antibody-protein mixture containing equimolar VEGF (Kangfang Bio, batch number: 20250926E), and SEB (final concentration of 50 ng / mL) were added to the culture plate containing the tumor cells. The culture plate was then returned to a 37°C incubator for further incubation for 24 / 72 hours.

[0221] Twenty-four hours later, the cell culture supernatant from the culture plate was collected, and human IL-2 was detected using enzyme-linked immunosorbent assay (ELISA). Seventy-two hours later, the cell culture supernatant from the culture plate was collected, and the secretion levels of human interferon-γ (IFNγ) and TNF-α were detected using enzyme-linked immunosorbent assay (ELISA).

[0222] The results are as follows Figures 1A to 1C , Figures 2A to 2C As shown in the results, in both colorectal cancer cell systems, both Ivonescimab monotherapy and VG2025 monotherapy could promote the secretion of IL-2, IFN-γ, and TNF-α by PBMCs; the combination therapy of Ivonescimab and VG2025 showed better activity than either Ivonescimab or VG2025 monotherapy, and in promoting IL-2 secretion, Ivonescimab and VG2025 in the combination therapy group had a significant synergistic effect.

[0223] Experimental Example 2: Clinical Trial of Liver Metastasis in Colorectal Cancer (CRC)

[0224] This is an open-label, multicenter, phase Ib / II clinical trial aimed at evaluating the safety and efficacy of Ivonescimab, oncolytic virus VG2025, with or without chemotherapy, for advanced CRC liver metastases. The study consists of a safety introductory phase and an expansion phase. Approximately 36-72 patients with unresectable CRC liver metastases who have failed first-line and / or second-line systemic antitumor therapy are planned for enrollment, with approximately 6-12 patients in the safety introductory phase and approximately 30-60 patients in the expansion phase.

[0225] Research Objective

[0226] Main purpose: To evaluate the safety and efficacy of Ivonescimab and VG2025 in combination with or without chemotherapy for the treatment of liver metastases of colorectal cancer (CRC).

[0227] Determine the recommended Phase II dose (RP2D) of Ivonescimab and VG2025 in combination with or without chemotherapy for the treatment of CRC liver metastases.

[0228] Secondary objective: To evaluate the pharmacokinetic (PK) characteristics of Ivonescimab in combination with VG2025 for the treatment of CRC liver metastases.

[0229] To evaluate the immunogenicity of Ivonescimab in combination with VG2025 for the treatment of CRC liver metastases.

[0230] Study endpoints

[0231] Primary endpoint: Safety: incidence and severity of dose-limiting toxicity (DLT) events, adverse events (AEs), clinically significant abnormal laboratory findings, vital signs, and electrocardiogram (ECG) results (based on NCI CTCAE version 5.0).

[0232] Efficacy: Objective response rate (ORR) assessed by investigators based on RECIST v1.1.

[0233] Secondary endpoint: Efficacy: Progression-free survival (PFS), disease control rate (DCR), duration of response (DoR), time to response (TTR), and overall survival (OS) as assessed by investigators based on RECIST v1.1.

[0234] PK characteristics: serum drug concentrations of Ivonescimab in individual subjects at different time points after administration.

[0235] Immunogenicity: The number and percentage of subjects with detectable anti-Ivonescimab antibodies, and the level of anti-HSV antibodies.

[0236] Research Design

[0237] The safety introductory phase comprises two study cohorts, enrolling CRC liver metastases patients who have failed first-line and / or second-line standard therapy including fluorouracil, oxaliplatin, and irinotecan. Each cohort will enroll 3-6 participants using a "3+3" approach. All cohorts will use a 28-day treatment cycle, with a 28-day treatment-to-live (DLT) observation period for safety evaluation following the first dose in the first treatment cycle. DLT observation and evaluation will be completed first for participants in cohort 1, and then progressed to cohort 2 for evaluation.

[0238] After DLT observational assessment is completed in any or all cohorts during the safety introductory phase, expanded enrollment at the recommended dose can proceed to further evaluate safety and efficacy. The recommended dose will be determined based on the results of the safety introductory phase, entering the expansion phase at a dose no higher than the recommended Phase 2 dose (RP2D) from the safety introductory phase. The expansion phase will enroll 30-60 participants, randomly assigned to cohort A and cohort B in a 2:1 ratio, stratified by ECOG score (0 vs 1). Participants in cohort A will receive Ivonescimab in combination with VG2025, while participants in cohort B will receive Ivonescimab monotherapy. The expansion enrollment phase will further monitor and evaluate the study's safety data. Investigators and sponsors will conduct a comprehensive discussion, incorporating efficacy and pharmacokinetic data, and will make decisions at any time regarding continuing, revising, or discontinuing the study cohorts, based on the situation in each cohort.

[0239] The safe induction phase (Phase Ib) is divided into two cohorts, Cohort 1 and Cohort 2, with the following treatment protocols: Queue 1: VG2025 (2.0×10 7 PFU (intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, intravenous infusion [ivgtt], day 8, day 22), each cycle is 28 days.

[0240] Queue 2: VG2025 (1.0×10 8 PFU (intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, IV drip, day 8, day 22), each cycle lasting 28 days.

[0241] The extended phase (Phase II) was divided into two cohorts, Cohort A and Cohort B, which enrolled CRC liver metastases patients who had failed first-line and / or second-line standard therapy including fluorouracil, oxaliplatin, and irinotecan. The treatment regimen was as follows: Cohort A: VG2025 (RP2D, intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, ivgtt, day 8, day 22), with each cycle lasting 28 days.

[0242] Cohort B: Ivonescimab 20 mg / kg, IV drip, day 1, day 15, every 28 days as one cycle.

[0243] All subjects were treated until the following occurred: the investigator determined that there was no longer any clinical benefit (based on a comprehensive assessment by the investigator using RECIST v1.1 imaging and clinical condition), toxicity became intolerable, 24 months of treatment was completed, or other criteria for discontinuing treatment were met, whichever occurred first. Safety and survival follow-ups were conducted after the discontinuation of all study drug treatment. Safety follow-ups were conducted on day 30 ± 7 after the last treatment to assess post-treatment safety. Survival follow-ups were conducted every 90 ± 14 days after the safety visit to obtain survival information and collect information on subsequent cancer treatment after the study treatment ended, until the subject died, withdrew informed consent, lost follow-up, or the study ended, whichever occurred first.

[0244] Researchers assessed efficacy according to RECIST v1.1 criteria. For the cohort with a 3-week treatment cycle, tumor assessment was performed every 6 weeks (±7 days) within 54 weeks of enrollment and every 12 weeks (±7 days) thereafter. For the cohort with a 4-week treatment cycle, tumor assessment was performed every 8 weeks (±7 days) within 56 weeks of enrollment and every 12 weeks (±7 days) thereafter. Subjects who discontinued treatment for reasons other than disease progression will continue to be followed up on their disease status until they begin other anti-tumor treatments, experience disease progression, withdraw informed consent, die, or the study ends, whichever occurs first. After the first recorded disease progression, if the subject does not exhibit clinical instability, disease progression must be confirmed no later than 8 weeks after 4 weeks (iRECIST criteria). During the study, researchers will collect a series of blood samples for pharmacokinetic, immunogenicity, and exploratory studies.

[0245] Treatment plan

[0246] Treatment consisted of 28-day cycles, with a time window of ±3 days per cycle. Three days prior to each dose, participants underwent various assessments, including vital signs, physical examination, laboratory tests, and performance status evaluation, to evaluate the safety and tolerability of continued treatment. Based on current research data, VG2025 (4.0 × 10⁻⁶ doses per cycle) was considered for the safe introduction phase. 7 PFU Q4W or 2.0×10 8VG2025 (PFU Q4W) in combination with Ivonescimab (20 mg / kg, Q2W) is indicated for treatment. Based on updated data from previous studies and safety and efficacy data from the safety rollout phase, the VG2025 dosage level may be adjusted. Investigators and sponsors will discuss and determine the RP2D and dosing frequency of VG2025 and Ivonescimab during the expanded enrollment phase, or other treatment regimens of VG2025 in combination with Ivonescimab.

[0247] The safe induction phase (Phase Ib) is divided into two cohorts, Cohort 1 and Cohort 2, with the following treatment protocols: Queue 1: VG2025 (2.0×10 7 PFU (intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, intravenous infusion [ivgtt], day 8, day 22), each cycle is 28 days.

[0248] Queue 2: VG2025 (1.0×10 8 PFU (intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, IV drip, day 8, day 22), each cycle lasting 28 days.

[0249] The extended phase (Phase II) was divided into two cohorts, Cohort A and Cohort B, which enrolled CRC liver metastases patients who had failed first-line and / or second-line standard therapy including fluorouracil, oxaliplatin, and irinotecan. The treatment regimen was as follows: Cohort A: VG2025 (RP2D, intratumoral injection, day 1, day 3) + Ivonescimab (20 mg / kg, ivgtt, day 8, day 22), with each cycle lasting 28 days.

[0250] Cohort B: Ivonescimab 20 mg / kg, IV drip, day 1, day 15, every 28 days as one cycle.

[0251] Treatment effect

[0252] The combination therapy of the anti-PD-1-anti-VEGFA bispecific antibody Ivonescimab and oncolytic virus VG2025 in patients with colorectal cancer liver metastases showed good safety and tolerability, and demonstrated good efficacy in treating colorectal cancer liver metastases.

[0253] Statistical analysis: The effectiveness analysis is based on an evaluable effectiveness analysis set and a security analysis set. The security analysis is based on the security analysis set.

[0254] The evaluable efficacy analysis set (EES) includes all participants who have received at least one treatment with the study drug and have measurable lesions at baseline (as defined by RECIST v1.1). The evaluable efficacy analysis set is used only for ORR, DCR, TTR, and DoR analyses.

[0255] The DLT evaluable set includes participants who were enrolled in the safety induction phase and developed DLT within 28 days of the first dose, or participants who did not develop DLT but received at least 80% of the expected dose of VG2025 and Ivonescimab as directed for the first 28 days and completed the safety follow-up through the 28-day DLT evaluation period.

[0256] The safety analysis set included participants who had received the study drug at least once.

[0257] The pharmacokinetic analysis set (PKS) includes all participants who have received at least one dose of Ivonescimab and have at least one effective post-dose plasma concentration data. The immunogenicity analysis set (IMS) includes all participants who have received at least one dose of Ivonescimab and have pre-dose and at least one post-dose immunogenicity assessment data.

[0258] Validity analysis: The primary endpoint of this study was the objective response rate (ORR) assessed according to RECIST v1.1. ORR and DCR analyses were performed on the evaluable validity set, with 95% exact two-sided confidence intervals estimated using the Clopper-Pearson method. DoR and TTR analyses were performed on the evaluable validity set, while PFS and OS analyses were performed on the safety set. For DoR, PFS, and OS, the median time and 95% CI were estimated using the Kaplan-Meier method, and Kaplan-Meier curves were plotted. Spider plots (for the change in the sum of target lesion diameters relative to baseline at each time point), waterfall plots (for the best percentage change in the sum of target lesion diameters relative to baseline), and swimlane plots were plotted to reflect the participants' tumor status during the study.

[0259] Security Analysis: Safety data includes adverse events, laboratory tests, and vital signs. All adverse events, investigational drug-related adverse events, and serious adverse events (SAEs) will be summarized, as well as AEs and AESIs leading to drug discontinuation. AEs will be graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) version 5.0 and coded using the Medical Dictionary for Regulatory Activities (MedDRA). Laboratory abnormalities and toxicity levels will be summarized according to NCI CTCAE version 5.0. The crosstabs for clinical laboratory tests will show the baseline CTCAE level and the change in the most severe level since baseline, and will list CTCAE level 3 or 4 clinical laboratory abnormalities.

[0260] Although specific embodiments of the invention have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details based on all the teachings disclosed, and all such changes are within the scope of protection of this invention. The full scope of this invention is given by the appended claims and any equivalents thereof.

Claims

1. The drug combination contains an effective amount of anti-PD-1-anti-VEGFA bispecific antibody and oncolytic virus, wherein: The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; Wherein, the first protein functional region is an immunoglobulin and the second protein functional region is a single-chain antibody; or, the first protein functional region is a single-chain antibody and the second protein functional region is an immunoglobulin. in, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; or, The immunoglobulin comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 7-9, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 10-12; and the single-chain antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences HCDR1-HCDR3 as shown in SEQ ID NOs: 1-3, and the light chain variable region comprises amino acid sequences LCDR1-LCDR3 as shown in SEQ ID NOs: 4-6; And, the oncolytic virus is herpes simplex virus type 1 (HSV-1).

2. The drug combination according to claim 1, wherein, The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 14; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. or, The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the immunoglobulin is shown in SEQ ID NO: 16; and the amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 13, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

14.

3. The pharmaceutical combination according to any one of claims 1 to 2, wherein: (1) The immunoglobulin is human IgG1 subtype; (2) The immunoglobulin is human IgG1 subtype, and according to the EU numbering system, the heavy chain constant region of the immunoglobulin has one of the following combinations of mutations: L234A and L235A; L234A and G237A; L235A and G237A; or L234A, L235A and G237A; (3) The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO: 18; (4) The first protein functional region is directly connected to the second protein functional region or is connected through a first linker; and / or the heavy chain variable region of the single-chain antibody is directly connected to the light chain variable region of the single-chain antibody or is connected through a second linker; (5) The first protein functional region is directly connected to the second protein functional region or is connected through a first linker; and / or the heavy chain variable region of the single chain antibody is directly connected to the light chain variable region of the single chain antibody or is connected through a second linker, and the first linker and the second linker are independently polypeptides of n peptides shown in SEQ ID NO: 19 in series, where n is 1, 2, 3, 4, 5 or 6; (6) The first protein functional region is directly connected to the second protein functional region or connected through a first linker; and / or the heavy chain variable region of the single-chain antibody is directly connected to the light chain variable region of the single-chain antibody or connected through a second linker; and the first linker and the second linker are independently polypeptides as shown in SEQ ID NO: 20; (7) The single-chain antibody is independently connected in sequence as follows: heavy chain variable region-linker-light chain variable region (VH-linker-VL), or light chain variable region-linker-heavy chain variable region (VL-linker-VH). (8) The amino acid sequence of the single-chain antibody is shown in SEQ ID NO: 21; (9) The first protein functional region and the second protein functional region are independently one, two or more; (10) The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; (11) The bispecific antibody is a bispecific antibody in the form of IgG-scFv; and / or (12) The bispecific antibody is a tetravalent bispecific antibody in the form of IgG-scFv.

4. The pharmaceutical combination according to any one of claims 1 to 3, wherein, The anti-PD-1-anti-VEGFA bispecific antibody is selected from the following 1)-7): 1） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; 2） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; 3） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; 4） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; 5） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; 6） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains are independently connected by one or more disulfide bonds; 7） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains.

5. The pharmaceutical combination according to any one of claims 1 to 4, wherein, The oncolytic virus is selected from: (1) Modified HSV-1; (2) Oncolytic virus VG2025; (3) Modified wild-type HSV-1; (4) Modified wild-type HSV-1 strain 17+; (5) Oncolytic viruses, whose ICP34.5 levels are lower in normal cells than in tumor cells; (6) Oncolytic viruses, whose ICP34.5 levels are lower in normal nerve cells than in tumor cells; (7) Modified HSV-1, and compared with wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the second copy is downregulated at the posttranscriptional mRNA level; (8) Modified HSV-1, in which the first copy of the two copies of the ICP34.5 gene is deleted or inactivated relative to wild-type HSV-1, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences. (9) Modified HSV-1, wherein, relative to wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences; the miRNA target sequences target miR-124 and / or miR-143; (10) Modified HSV-1, wherein, relative to wild-type HSV-1, the first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy of the ICP34.5 gene contains 2-10 miRNA target sequences; the miRNA target sequences target miR-124 and / or miR-143; the sequence of miR-124 is shown in SEQ ID NO: 30; the sequence of miR-143 is shown in SEQ ID NO: 31; (11) Modified HSV-1, and the natural promoter of ICP27 was replaced by a tumor-specific promoter; (12) Modified HSV-1, and the natural promoter of ICP27 is replaced by the human carcinoembryonic antigen promoter; (13) Modified HSV-1, and the natural promoter of ICP27 is replaced by the human carcinoembryonic antigen promoter; the sequence of the human carcinoembryonic antigen promoter is shown in SEQ ID NO: 23; (14) A modified HSV-1, and comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; (15) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. (16) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; the expression cassette being driven by a CXCR4 promoter; (17) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (18) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15 and IL-15Rα; the expression cassette being driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25; (19) A modified HSV-1, comprising an expression cassette expressing one or more of IL12, IL15, and IL-15Rα; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO:

25. (20) Modified HSV-1, and comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide. (21) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (22) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; (23) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. (24) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (25) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by the CXCR4 promoter; (26) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15 and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of IL12 is shown in SEQ ID NO: 26, the amino acid sequence of IL15 is shown in SEQ ID NO: 27, and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO: 25; (27) A modified HSV-1, comprising an expression cassette expressing IL-12, IL-15, and IL-15Rα in tandem, wherein the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide; wherein the amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; the amino acid sequence of IL12 is shown in SEQ ID NO: 26; the amino acid sequence of IL15 is shown in SEQ ID NO: 27; and the amino acid sequence of IL-15Rα is shown in SEQ ID NO: 29; the expression cassette is driven by a CXCR4 promoter; the sequence of the CXCR4 promoter is shown in SEQ ID NO:

25. (28) Modified HSV-1, and the C-terminal amino acid positions 877-904 of glycoprotein B were truncated; (29) Modified HSV-1, and the C-terminal amino acid positions 877-904 of glycoprotein B were truncated; the amino acid sequence of the truncated glycoprotein B is shown in SEQ ID NO: 24; (30) Modified wild-type HSV-1 or modified wild-type HSV-1 strain 17+, and possessing any one, two, three, or all four of the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; (31) Any one or any combination of the above items.

6. The pharmaceutical combination according to any one of claims 1 to 5, further comprising an effective amount of a chemotherapy drug, wherein, The chemotherapy drugs are those used in the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen.

7. The pharmaceutical combination according to any one of claims 1 to 6, comprising an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody and an oncolytic virus, or comprising an effective amount of an anti-PD-1-anti-VEGFA bispecific antibody, an oncolytic virus and a chemotherapeutic agent, and selected from any one of (1) to (15) below: （1） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; （2） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; （3） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; （4） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; （5） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; （6） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; （7） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; （8） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; （9） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; （10） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drugs are chemotherapy drugs of the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; （11） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; （12） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain variable region of the immunoglobulin is shown in SEQ ID NO: 13, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO:

14. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; （13） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; （14） The anti-PD-1-anti-VEGFA bispecific antibody includes: Targeting the first protein functional region of VEGFA, and Targeting the second protein functional region of PD-1; The first protein has one functional region, and the second protein has two functional regions; Wherein, the first protein functional region is an immunoglobulin, and the second protein functional region is a single-chain antibody; The amino acid sequence of the heavy chain of the immunoglobulin is shown in SEQ ID NO: 17, and the amino acid sequence of its light chain is shown in SEQ ID NO:

18. The amino acid sequence of the heavy chain variable region of the single-chain antibody is shown in SEQ ID NO: 15, and the amino acid sequence of the light chain variable region of the single-chain antibody is shown in SEQ ID NO:

16. The single-chain antibody is attached to the C-terminus or N-terminus of the heavy chain of the immunoglobulin; The first protein functional region and the second protein functional region are connected by a first linker; and the heavy chain variable region of the single-chain antibody and the light chain variable region of the single-chain antibody are connected by a second linker; the first linker and the second linker may be the same or different; The amino acid sequences of both the first and second linkers are shown in SEQ ID NO: 20; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine; （15） The anti-PD-1-anti-VEGFA bispecific antibody is a tetramer formed by two identical first peptide chains and two identical second peptide chains, wherein... The amino acid sequence of the first peptide chain is shown in SEQ ID NO: 22; and the amino acid sequence of the second peptide chain is shown in SEQ ID NO: 18; The first peptide chain and the second peptide chain, and / or the two first peptide chains, are independently connected by 1, 2, 3, or 4 disulfide bonds; and There are no disulfide bonds connecting the two second peptide chains; The oncolytic virus is a modified wild-type HSV-1 strain 17+, and has the following characteristics: (i) The first copy of the two copies of the ICP34.5 gene is deleted or inactivated, and the 3'UTR region of the second copy contains binding sites for miR-124 and miR-143, wherein, The sequence of miR-124 is shown in SEQ ID NO: 30, and The sequence of miR-143 is shown in SEQ ID NO: 31; (ii) The natural promoter of ICP27 is replaced by the human carcinoembryonic antigen (CEA) promoter; and the sequence of the CEA promoter is shown in SEQ ID NO: 23; (iii) Contains an expression cassette expressing IL12, IL15, and the IL15 receptor α subunit, the expression cassette being driven by the CXCR4 promoter, wherein IL-12, IL-15, and IL-15Rα are tandemly linked, and the N-terminus of IL-15 and IL-15Rα are respectively linked to a 2A self-cleaving peptide, wherein... The amino acid sequence of IL12 is shown in SEQ ID NO:

26. The amino acid sequence of IL15 is shown in SEQ ID NO:

27. The amino acid sequence of IL-15Rα is shown in SEQ ID NO:

29. The sequence of the CXCR4 promoter is shown in SEQ ID NO: 25, and The amino acid sequence of the 2A self-cleaving peptide is shown in SEQ ID NO: 28; (iv) The amino acid sequence of glycoprotein B is shown in SEQ ID NO: 24; The chemotherapy drug is: Oxaliplatin, leucovorin calcium, and 5-fluorouracil Irinotecan, leucovorin calcium, and 5-fluorouracil, or Oxaliplatin and capecitabine.

8. The pharmaceutical combination according to any one of claims 1 to 7, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody is mixed with an oncolytic virus, and the drug combination is a pharmaceutical composition; or The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are packaged in separate containers, and the drug combination is a combination product. (2) Based on the mass of the anti-PD-1-anti-VEGFA bispecific antibody, the unit dose of the anti-PD-1-anti-VEGFA bispecific antibody is 100mg-2500mg, 100mg-2000mg, 100mg-1500mg, 100mg-1200mg, 100mg-1000mg, 200mg-800mg, 200mg-500mg, 300mg-600mg, 400mg-500mg, or 450mg; (3) The titer of the oncolytic virus is approximately 5 × 10⁻⁶. 4 pfu / mL - approximately 2 × 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 10 9 pfu / mL, approximately 10 6 pfu / mL - approximately 10 7 pfu / mL, approximately 10 7 pfu / mL - approximately 10 8 pfu / mL, approximately 10 8 pfu / mL - approximately 10 9 pfu / mL or approximately 10 8 pfu / mL; (4) The drug combination further comprises one or more pharmaceutically acceptable excipients; (5) The drug combination also includes a product instruction manual.

9. The pharmaceutical combination according to any one of claims 1 to 8, for the treatment or prevention of tumors.

10. The pharmaceutical combination according to claim 9, wherein, The tumor is selected from one or more of the following: Colorectal cancer, liver cancer, advanced tumors, metastatic tumors, colorectal cancer liver metastases, advanced colorectal cancer liver metastases, hepatocellular carcinoma, advanced liver cancer, and advanced hepatocellular carcinoma.

11. The pharmaceutical combination according to any one of claims 1 to 10, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Alternatively, the anti-PD-1-anti-VEGFA bispecific antibody may be administered after the oncolytic virus, with an interval of 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg or 1200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, and no administration is given during the rest of the cycle. Each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or oral administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) A combination of any of the items selected from (1)-(8) above; (10) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. (11) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion; (12) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. or (13) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, by intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion.

12. Use of the pharmaceutical combination according to any one of claims 1 to 8 in the preparation of a medicament for treating or preventing tumors.

13. The use according to claim 12, wherein, The tumor is selected from one or more of the following: Colorectal cancer, liver cancer, advanced tumors, metastatic tumors, colorectal cancer liver metastases, advanced colorectal cancer liver metastases, hepatocellular carcinoma, advanced liver cancer, and advanced hepatocellular carcinoma.

14. The use according to any one of claims 12 to 13, wherein: (1) The anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are administered independently before or after surgery, and / or before or after radiotherapy; (2) The anti-PD-1-anti-VEGFA bispecific antibody is administered before, simultaneously with, or after the oncolytic virus; Alternatively, the anti-PD-1-anti-VEGFA bispecific antibody may be administered after the oncolytic virus, with an interval of 3, 4, 5, 6 or 7 days. (3) The single dose of the anti-PD-1-anti-VEGFA bispecific antibody is 0.1-100 mg, 5-50 mg, 1-10 mg, 5-15 mg, 15-25 mg or 20 mg per kilogram of body weight; or 10-2000 mg, 50-1500 mg, 100-1400 mg, 150-1300 mg, 500-1500 mg or 1200 mg per subject; (4) The anti-PD-1-anti-VEGFA bispecific antibody is administered once every half day, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks; (5) The single dose of the oncolytic virus is approximately 5 × 10⁻⁶ per subject. 4 pfu to approximately 2 × 10 9 pfu, approximately 10 6 pfu to approximately 10 9 pfu, approximately 10 6 pfu to approximately 10 7 pfu, approximately 10 7 pfu to approximately 10 8 PFU or about 10 8 pfu to approximately 10 9 pfu; (6) The oncolytic virus is administered according to a cycle. The beginning of each cycle is two or three consecutive days of administration once a day, and no administration is given during the rest of the cycle. Each cycle consists of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. (7) The administration methods of the anti-PD-1-anti-VEGFA bispecific antibody and the oncolytic virus are independently intratumoral injection, intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or oral administration; (8) The chemotherapy drugs are administered according to the mFOLFOX6 regimen, FOLFIRI regimen, or CapeOX regimen; (9) A combination of any of the items selected from (1)-(8) above; (10) The oncolytic virus was administered once daily on days 1-2 or 1-3, or once on day 1 and once on day 3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, or once on day 1 and once on day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. (11) The oncolytic virus was administered once daily on days 1-2 or 1-3, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; The anti-PD-1-anti-VEGFA bispecific antibody was administered once on day 8 and once on day 22, at a dose of 20 mg per kilogram of body weight, via intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion; (12) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, via intravenous infusion; and A cycle is defined as either 21 days or 28 days. or (13) VG2025 was administered once daily on days 1-2 or 1-3, or once on days 1 and 3 respectively, at a dose of 2.0 × 10⁻⁶ per subject. 7 PFU up to 1.0×10 8 PFU, intratumoral injection; Ivoxis was administered once on day 8 and day 22, or once on day 1 and day 15, at a dose of 20 mg per kilogram of body weight, by intravenous infusion. A cycle is defined as either 21 days or 28 days. Furthermore, chemotherapy drugs were administered once on day 8 and once on day 22, according to the mFOLFOX6, FOLFIRI, or CapeOX regimens, via intravenous infusion.

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