Application of oncolytic virus combined targeting antibody and chemotherapeutic drug in inhibition of liver metastasis of colon cancer

Abstract

The invention discloses an application of an oncolytic virus, a VEGF / EGF targeting antibody and a chemotherapeutic drug in synergistic treatment of liver metastasis of colon cancer. Specifically, the invention provides an application of an active component combination, the active component combination comprises a recombinant human adenovirus type 5, a VEGF / EGF targeting antibody and a chemotherapeutic drug, and the combination is used for preparing a pharmaceutical composition for synergistically treating tumors. Clinical experiments prove that the active component composition disclosed by the invention can effectively and synergistically inhibit liver metastasis of colon cancer and does not generate side effects.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-tumor drugs, and specifically to the use of oncolytic viruses combined with VEGF / EGF targeting antibodies and chemotherapy drugs in inhibiting liver metastasis of colon cancer. Background Art

[0002] Colon cancer is one of the most common digestive tract malignancies in humans. The incidence and mortality of colorectal cancer (CRC) in my country have been on the rise. The 2018 China Cancer Statistics Report shows that the incidence and mortality of colorectal cancer in my country ranked third and fifth among all malignant tumors, with 376,000 new cases and 191,000 deaths. The incidence in cities is much higher than in rural areas, and the incidence of colon cancer has increased significantly. Most patients are already in the middle and late stages when diagnosed.

[0003] The basic treatment for colon cancer is surgical resection, with adjuvant chemotherapy for patients with high-risk stage II and stage III / IV colon cancer. Most early-stage colon patients can undergo successful surgical resection, but for stage II / III patients, recurrence often occurs, which is closely related to metastasis. About 30% of patients have varying degrees of liver metastasis when they are first diagnosed, and about 20% of patients will develop metachronous liver metastasis during tumor progression, of which the vast majority (80%-90%) of liver metastases cannot be radically removed initially.

[0004] Metastasis is the main cause of death in patients with colon cancer. Currently, the main treatment drugs are fluorouracil / leucovorin combined with oxaliplatin and / or irinotecan; chemical drugs combined with molecular targeted drugs VEGF / EGF targeted antibodies can further improve the efficacy. However, the toxicity and adverse reactions of chemotherapy and antibody resistance require a more optimized treatment plan. Summary of the invention

[0005] The purpose of the present invention is to provide an application of an active ingredient combination comprising an oncolytic virus, a VEGF / EGF targeting antibody and a chemotherapeutic drug in the treatment of liver metastasis of colon cancer, and a drug comprising the active ingredient combination.

[0006] The first aspect of the present invention provides a use of an active ingredient combination, the active ingredient combination comprising:

[0007] The first active ingredient is oncolytic virus, the second active ingredient is VEGF / EGF targeting antibody and the third active ingredient is chemotherapeutic drug, and the combination is used to prepare a pharmaceutical composition or a drug kit for synergistically treating liver metastasis of colon cancer.

[0008] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0009] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0010] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0011] In another preferred embodiment, the VEGF / EGF targeting antibody is selected from the following group: bevacizumab, cetuximab, or a combination thereof.

[0012] In another preferred embodiment, the second active ingredient is the VEGF targeting antibody bevacizumab.

[0013] In another preferred embodiment, the second active ingredient is the EGF targeting antibody cetuximab.

[0014] In another preferred embodiment, the chemotherapy drug is selected from the following group: irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

[0015] In another preferred embodiment, the third active ingredient is a chemotherapy drug combination comprising irinotecan, calcium folinate and fluorouracil.

[0016] In another preferred embodiment, the third active ingredient is a chemotherapy drug combination comprising oxaliplatin, calcium folinate and fluorouracil.

[0017] In another preferred embodiment, the third active ingredient is a chemotherapy drug combination comprising irinotecan, oxaliplatin, calcium folinate and fluorouracil.

[0018] In another preferred embodiment, the third active ingredient is a chemotherapy drug combination comprising oxaliplatin and S-1.

[0019] In another preferred embodiment, the third active ingredient is a chemotherapy drug combination comprising oxaliplatin and capecitabine.

[0020] In another preferred embodiment, the third active ingredient comprises capecitabine.

[0021] In another preferred embodiment, the oncolytic virus is administered at a rate of 0.5×10 12 VP to 2.5×10 12 VP is administered at a dose of 1 to 20 mg / kg, preferably 2 to 15 mg / kg, and more preferably 3 to 10 mg / kg.

[0022] The second aspect of the present invention provides a combination of active ingredients, which consists of an oncolytic virus, a VEGF / EGF targeting antibody and a chemotherapeutic drug.

[0023] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0024] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0025] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0026] In another preferred embodiment, the VEGF / EGF targeting antibody is selected from the following group: bevacizumab, cetuximab, or a combination thereof.

[0027] In another preferred embodiment, the chemotherapy drug is selected from the following group: irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

[0028] In another preferred embodiment, the active ingredient combination consists of recombinant human adenovirus type 5, bevacizumab, and chemotherapeutic drugs.

[0029] In another preferred embodiment, the chemotherapy drug is a combination of irinotecan, calcium folinate and fluorouracil.

[0030] In another preferred embodiment, the chemotherapy drug is capecitabine.

[0031] In another preferred embodiment, the chemotherapy drug is a combination of irinotecan, oxaliplatin, calcium folinate and fluorouracil.

[0032] In another preferred embodiment, the chemotherapy drug is a combination of oxaliplatin and capecitabine.

[0033] In another preferred embodiment, the active ingredient combination consists of recombinant human adenovirus type 5, cetuximab, and chemotherapy drugs.

[0034] In another preferred embodiment, the chemotherapy drug is a combination of oxaliplatin, calcium folinate and fluorouracil.

[0035] In another preferred embodiment, the chemotherapy drug is a combination of oxaliplatin and S-1.

[0036] In another preferred embodiment, the chemotherapy drug is a combination of irinotecan, oxaliplatin, calcium folinate and fluorouracil.

[0037] The third aspect of the present invention provides a pharmaceutical composition for treating liver metastasis of colon cancer, wherein the pharmaceutical composition comprises (a) an oncolytic virus, (b) a VEGF / EGF targeting antibody, (c) a chemotherapeutic drug, and (d) a pharmaceutically acceptable carrier.

[0038] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0039] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0040] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0041] In another preferred embodiment, the VEGF / EGF targeting antibody is selected from the following group: bevacizumab, cetuximab, or a combination thereof.

[0042] In another preferred embodiment, the chemotherapy drug is selected from the following group: irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

[0043] A fourth aspect of the present invention provides a medicine kit, comprising:

[0044] (a) a first preparation, wherein the first preparation contains an oncolytic virus and a pharmaceutically acceptable carrier;

[0045] (b) a second formulation comprising a VEGF / EGF targeting antibody and a pharmaceutically acceptable carrier;

[0046] (c) a third preparation comprising a chemotherapeutic drug and a pharmaceutically acceptable carrier; and

[0047] (d) instructions describing a method for treating colon cancer liver metastasis by combining an oncolytic virus, a VEGF / EGF targeting antibody, and a chemotherapeutic drug.

[0048] In another preferred embodiment, the first preparation, the second preparation and the third preparation are independent of each other.

[0049] In another preferred embodiment, the first preparation, the second preparation and the third preparation are freeze-dried preparations or liquid preparations.

[0050] In another preferred embodiment, the first preparation, the second preparation, and the third preparation are injections.

[0051] In another preferred embodiment, the first preparation is administered before the second preparation and the third preparation.

[0052] In another preferred embodiment, the second preparation and the third preparation are administered simultaneously.

[0053] In another preferred embodiment, the second preparation is administered before the third preparation.

[0054] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0055] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0056] In another preferred embodiment, the VEGF / EGF targeting antibody is selected from the following group: bevacizumab, cetuximab, or a combination thereof.

[0057] In another preferred embodiment, the chemotherapy drug is selected from the following group: irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

[0058] The fifth aspect of the present invention provides a method for treating liver metastasis of colon cancer, which comprises administering a therapeutically effective amount of the active ingredient combination as described in the second aspect of the present invention and / or the pharmaceutical composition as described in the third aspect of the present invention to a subject in need, thereby treating liver metastasis of colon cancer.

[0059] In another preferred embodiment, the subject in need is a human or a non-human mammal.

[0060] In another preferred embodiment, the non-human mammals include monkeys, gorillas, cows, pigs, dogs, sheep, rabbits or mice.

[0061] In another preferred embodiment, the subject in need thereof suffers from colon cancer.

[0062] In another preferred embodiment, the subject in need thereof suffers from colon cancer with liver metastasis.

[0063] In another preferred embodiment, in the active ingredient combination or pharmaceutical composition, the oncolytic virus is administered by intratumoral injection; the VEGF / EGF targeting antibody is administered by intravenous injection; and the chemotherapy drug is administered by intravenous injection, intraperitoneal injection, intravenous infusion or oral administration.

[0064] In another preferred embodiment, the method further comprises administering supportive therapeutic drugs for treating tumors to a subject in need thereof.

[0065] In another preferred embodiment, the supportive treatment drugs include: antibiotics, analgesics, nutritional drugs or a combination thereof.

[0066] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 The tumor size before and after treatment of a case in clinical trial 1 using recombinant human adenovirus type 5 in combination with VEGF-targeting antibodies and chemotherapy drugs is shown.

[0068] Figure 2 The tumor size before and after treatment of a case in clinical trial 2 using recombinant human adenovirus type 5 in combination with VEGF-targeting antibodies and chemotherapy drugs is shown.

[0069] Figure 3 The tumor size before and after treatment of a case in clinical trial 3 using recombinant human adenovirus type 5 in combination with VEGF-targeting antibodies and chemotherapy drugs is shown.

[0070] Figure 4 The tumor size before and after treatment of a case from clinical trial 4 using a recombinant human adenovirus type 5 in combination with a VEGF-targeting antibody and chemotherapy drugs is shown.

[0071] Figure 5 The tumor size before and after treatment of a case in clinical trial 1 using a recombinant human adenovirus type 5 in combination with an EGF-targeting antibody and chemotherapy drugs is shown.

[0072] Figure 6 The tumor size before and after treatment of a case in clinical trial 2 using a recombinant human adenovirus type 5 in combination with an EGF-targeting antibody and chemotherapy drugs is shown.

[0073] Figure 7 The tumor size before and after treatment of a case in clinical trial 3 using a recombinant human adenovirus type 5 in combination with an EGF-targeting antibody and chemotherapy drugs is shown.

[0074] Figure 8 The tumor size before and after treatment of a case in clinical trial 4 using a recombinant human adenovirus type 5 in combination with an EGF-targeting antibody and chemotherapy drugs is shown. DETAILED DESCRIPTION

[0075] After extensive and in-depth research, the inventor unexpectedly discovered for the first time that when a specific recombinant human adenovirus type 5 (Ankorui) is used in combination with VEGF / EGF targeting antibodies and chemotherapy drugs, it has a synergistic therapeutic effect in inhibiting liver metastasis of colon cancer. Clinical experiments have shown that the combined use of recombinant human adenovirus type 5 with VEGF / EGF targeting antibodies and chemotherapy drugs can effectively inhibit the growth of metastatic tumors. On this basis, the present invention was completed.

[0076] the term

[0077] As used herein, the term "first active ingredient" refers to recombinant human adenovirus type 5, or oncolytic virus. It should be understood that the term includes derivatives having the same function as recombinant human adenovirus type 5. In a preferred embodiment of the present invention, the "first active ingredient" is a recombinant human adenovirus type 5 injection with the trade name of Ankorui.

[0078] As used herein, the term "second active ingredient" refers to a VEGF / EGF targeting antibody. In a preferred embodiment of the present invention, the "second active ingredient" is a VEGF targeting antibody bevacizumab. In another preferred embodiment of the present invention, the "second active ingredient" is an EGF targeting antibody cetuximab.

[0079] As used herein, the term "third active ingredient" refers to a chemotherapeutic drug. The chemotherapeutic drug is selected from the group consisting of irinotecan, oxaliplatin, folinic acid, fluorouracil, S-1, capecitabine, or a combination thereof. In a preferred embodiment of the present invention, the "third active ingredient" refers to a chemotherapeutic drug combination comprising irinotecan, folinic acid and fluorouracil (i.e., FOLFIRI). In another preferred embodiment of the present invention, the "third active ingredient" refers to a chemotherapeutic drug combination comprising oxaliplatin (Aiheng or Eloxadin), folinic acid and fluorouracil (i.e., FOLFOX). In another preferred embodiment of the present invention, the "third active ingredient" refers to a chemotherapeutic drug combination comprising irinotecan, oxaliplatin (Aiheng or Eloxadin), folinic acid and fluorouracil (i.e., FOLFOXIRI). In another preferred embodiment of the present invention, the "third active ingredient" refers to a chemotherapeutic drug combination comprising oxaliplatin (Aiheng or Eloxadin) and S-1 (i.e., SOX). In another preferred embodiment of the present invention, the "third active ingredient" refers to a chemotherapy drug combination comprising oxaliplatin (Aixanthin or Eloxatin) and capecitabine (i.e., XELOX). In another preferred embodiment of the present invention, the "third active ingredient" comprises the chemotherapy drug capecitabine.

[0080] As used herein, the terms "active ingredient combination of the present invention", "substance combination of the present invention" and "pharmaceutical ingredient combination of the present invention" are used interchangeably to refer to the combination of the above-mentioned first active ingredient, second active ingredient and third active ingredient.

[0081] As used herein, the term "pharmaceutical composition of the present invention" refers to a composition comprising or containing a first active ingredient (or a first preparation containing the first active ingredient), a second active ingredient (or a second preparation containing the second active ingredient), and a third active ingredient (or a third preparation containing the third active ingredient). In the present invention, the first preparation, the second preparation, and the third preparation may be the same or different preparations. In addition, the first preparation, the second preparation, and the third preparation may be the same preparation, or each independent preparation.

[0082] Recombinant human adenovirus type 5 injection

[0083] The recombinant human adenovirus type 5 injection used in the present invention, with the trade name of Ankorui, is the first oncolytic virus therapeutic drug approved by the China Food and Drug Administration and launched on the market in 2006. It is an oncolytic adenovirus obtained by genetically recombining human adenovirus type 5 (Ad5) using genetic engineering technology. It mainly deletes the E1B-55kDa (i.e., 55kD in the E1B region of type 5 adenovirus, encoded by the E1B-55k gene) and E3 region gene fragments of human adenovirus type 5, and has the characteristic of specific replication in tumor cells, which ultimately leads to oncolysis.

[0084] The oncolytic mechanism of oncolytic viruses is constantly updated with the progress of tumor immunity research. It was previously believed that because some viruses are naturally tropistic for tumors, they can selectively infect tumor cells and lyse tumor cells to kill the tumor. As the research continues to deepen, it has been found that oncolytic viruses not only have the ability to directly lyse tumors, but also can improve the tumor microenvironment in an immunosuppressive state, turning "cold" tumors into "hot" tumors, thereby activating innate immunity and adaptive immunity, so that the body's immune system that was originally evaded by the tumor is mobilized to attack the tumor.

[0085] Adenovirus E1B-55kDa and E3 region gene fragments

[0086] Adenovirus is a type of pathogen that has been studied in depth. Since the 1960s, adenovirus has been used as a gene expression vector and has been widely used in the field of molecular biology. It is the most commonly used viral vector for gene therapy. Adenovirus vectors have many characteristics that other vectors do not have. Various types of adenovirus vectors have been successfully constructed and applied in engineering.

[0087] Adenovirus is a DNA virus without an envelope. It is composed of 252 capsomers arranged in an icosahedral pattern and is a particle of 70 to 90 nm. The capsid contains a linear double-stranded DNA molecule with an inverted repeat sequence of about 100 bp at each end. Since the 5' end of each DNA chain has a relative molecular mass of 55×10 3 The protein molecules of Da are covalently bound to form a circular structure of double-stranded DNA.

[0088] The adenovirus E3 region gene is an early transcription unit of the adenovirus genome. Its promoter is activated by the E1A protein and then transcribed into protein. The E3 protein mainly destroys the host's immune defense mechanism, protecting the adenovirus from attack by the immune system in the host.

[0089] The recombinant human adenovirus type 5 used in the present invention has been modified, and after the E3 region gene is deleted, it only has the ability to replicate in the host cell without damaging the host's immune system.

[0090] VEGF / EGF Targeting Antibodies

[0091] Molecular targeted therapy for tumors refers to the use of the biological differences between tumor cells and normal cell molecules, taking the tumor's proto-oncogene products or their signal transduction pathways as therapeutic targets, and blocking the signal transduction pathways through monoclonal antibodies or enzyme inhibitors, thereby achieving the purpose of inhibiting tumor growth. Molecular targeted therapy drugs are divided into monoclonal antibodies targeting specific cell markers, signal transduction inhibitors, anti-angiogenic drugs, and drugs targeting certain cytogenetic markers or oncogene products.

[0092] Bevacizumab (trade name Avastin, Ankeda) is a recombinant humanized immunoglobulin G1 (IgG1) monoclonal antibody that can bind to VEGF-A and inhibit its binding to VEGF receptor-2 (VEGFR-2), thereby inhibiting the biological effects of VEGF, including affecting vascular permeability, proliferation, and endothelial cell migration and survival, thereby inhibiting tumor angiogenesis, growth, and metastasis. Bevacizumab is currently used to treat various types of metastatic cancer. Its combination with fluoropyrimidine-based chemotherapy is suitable for the treatment of patients with metastatic colorectal cancer.

[0093] Cetuximab, also known as Erbitux (IMC-C225, Erbitux), is an IgG1 monoclonal antibody against EGF receptor. After the two specifically bind, it inhibits the tyrosine kinase (TK) after binding to the EGF receptor, blocks the intracellular signal transduction pathway, thereby inhibiting the proliferation of cancer cells, inducing apoptosis of cancer cells, and reducing the production of matrix metalloproteinases and vascular endothelial growth factor. It is suitable for the treatment of patients with metastatic colorectal cancer in combination with fluoropyrimidine-based chemotherapy.

[0094] The chemotherapeutic drug of the present invention

[0095] The present invention provides chemotherapeutic drugs for treating liver metastasis of colon cancer, wherein the chemotherapeutic drugs include (but are not limited to): irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

[0096] In a preferred embodiment of the present invention, the chemotherapy drug is a chemotherapy drug combination of irinotecan, calcium folinate and fluorouracil.

[0097] In a preferred embodiment of the present invention, the chemotherapy drug is a chemotherapy drug combination of oxaliplatin, calcium folinate and fluorouracil.

[0098] In a preferred embodiment of the present invention, the chemotherapy drug is a chemotherapy drug combination of irinotecan, oxaliplatin, calcium folinate and fluorouracil.

[0099] In a preferred embodiment of the present invention, the chemotherapy drug is a chemotherapy drug combination of oxaliplatin and S-1.

[0100] In a preferred embodiment of the present invention, the chemotherapy drug is a chemotherapy drug combination of oxaliplatin and capecitabine.

[0101] In a preferred embodiment of the present invention, the chemotherapy drug is capecitabine.

[0102] Pharmaceutical composition and administration method

[0103] The present invention also provides a composition which can be used for synergistically treating liver metastasis of colon cancer.

[0104] The pharmaceutical composition of the present invention comprises: an effective amount of recombinant human adenovirus type 5, an effective amount of VEGF targeting antibody, an effective amount of chemotherapeutic drug, and a pharmaceutically acceptable carrier.

[0105] Generally, the recombinant human adenovirus type 5, or VEGF targeting antibody, or chemotherapeutic drug of the present invention can be formulated in a non-toxic, inert and pharmaceutically acceptable carrier medium, wherein the pH is generally about 5-8, preferably, about 6-8.

[0106] As used herein, "pharmaceutically acceptable carrier" refers to a carrier used for administration of therapeutic agents, including various excipients and diluents.

[0107] The term refers to pharmaceutical carriers that are not essential active ingredients and are not overly toxic after administration. Suitable carriers are well known to those of ordinary skill in the art. Pharmaceutically acceptable carriers in the composition may contain liquids such as water, saline, and buffers. In addition, auxiliary substances such as fillers, lubricants, glidants, wetting agents or emulsifiers, pH buffer substances, etc. may also be present in these carriers. The carriers may also contain cell transfection reagents.

[0108] As used herein, the term "effective amount" or "effective dose" refers to an amount that can produce a function or activity on humans and / or animals and / or cells and can be accepted by humans and / or animals.

[0109] As used herein, "pharmaceutically acceptable" ingredients are suitable for use in humans and / or mammals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier used for the administration of therapeutic agents, including various excipients and diluents. Such carriers include (but are not limited to): saline, buffer, glucose, water, glycerol, polysorbate, ethanol, and combinations thereof. Generally, the pharmaceutical preparation should match the mode of administration. The pharmaceutical composition of the present invention can be prepared in the form of an injection, for example, by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants. The pharmaceutical composition is preferably manufactured under sterile conditions. The amount of active ingredient administered is a therapeutically effective amount. The pharmaceutical preparation of the present invention can also be prepared as a sustained-release preparation.

[0110] Furthermore, the active ingredient combination of the present invention may also be used together with other therapeutic agents (such as anti-tumor agents or chemotherapeutic agents).

[0111] When using a pharmaceutical composition, a safe and effective amount of a combination of active ingredients (including a first active ingredient (or its preparation), a second active ingredient (or its preparation) and a third active ingredient (or its preparation)) is administered to a mammal.

[0112] It should be understood that the effective amount of the first active ingredient (or its preparation) and / or the second active ingredient (or its preparation) and / or the third active ingredient (or its preparation) in the active ingredient combination of the present invention may vary with the mode of administration and the severity of the tumor. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters such as bioavailability, metabolism, half-life, etc.; the severity of the tumor, the patient's weight, the patient's immune status, the route of administration, etc.

[0113] Typically, for the first active ingredient, the therapeutically effective amount is usually at least 0.5×10 12 VP to 2.5×10 12 VP, of course, the specific dose should also take into account factors such as tumor volume and patient health status, which are all within the skills of a skilled physician.

[0114] For the second active ingredient, its safe and effective amount is usually at least about 1 mg / kg body weight, and in most cases does not exceed about 20 mg / kg body weight, preferably the dose is about 2 mg / kg body weight to 10 mg / kg body weight. Of course, the specific dose should also take into account factors such as the type of drug used, dosage form, route of administration, and patient health status, which are all within the skill range of skilled physicians.

[0115] There is no particular limitation on the administration method of the pharmaceutical composition of the present invention. Representative examples include (but are not limited to): intratumor injection, intraperitoneal injection, intravenous injection, intravenous infusion, and oral administration.

[0116] Pill Box

[0117] The present invention provides a medicine box, comprising:

[0118] Component (1): a preparation containing recombinant human adenovirus type 5;

[0119] Component (2): a preparation comprising a VEGF / EGF targeting antibody;

[0120] Component (3): a preparation containing a chemotherapeutic drug; and

[0121] Component (4): instructions.

[0122] The preparation containing recombinant human adenovirus type 5 includes (but is not limited to): a lyophilized preparation, a liquid preparation or an injection.

[0123] The preparation containing the VEGF / EGF targeting antibody includes (but is not limited to): a lyophilized preparation, a liquid preparation, or an injection.

[0124] The preparation containing the chemotherapy drug includes (but is not limited to): a lyophilized preparation, a liquid preparation, or an injection.

[0125] Typically, the kit contains one or more (e.g., at least two) unit dosage forms containing recombinant human adenovirus type 5 and one or more (e.g., at least two) unit dosage forms containing VEGF / EGF targeting antibodies, and one or more (e.g., at least two) unit dosage forms containing chemotherapeutic drugs; preferably, 4-10 of each.

[0126] As used herein, the term "unit dosage form" means that for the convenience of administration, a composition is prepared into a dosage form required for a single administration, including but not limited to various solid doses (such as tablets), liquid doses, capsules, and sustained-release preparations.

[0127] In the present invention, the unit dosage form of the recombinant human adenovirus type 5 is usually a liquid dosage form.

[0128] The instructions provided by the present invention may include the following description: the method of using the drug kit is to simultaneously use a unit dosage form containing a recombinant human adenovirus type 5, a unit dosage form containing a VEGF / EGF targeting antibody, and a unit dosage form containing a chemotherapeutic drug.

[0129] The medicine kit provided by the present invention is prepared by the following steps: placing a preparation containing recombinant human adenovirus type 5, a preparation containing VEGF / EGF targeting antibody, a preparation containing chemotherapy drugs and instructions together to form a medicine kit.

[0130] The preparation containing recombinant human adenovirus type 5 preferably contains a unit dosage form of recombinant human adenovirus type 5, the preparation containing VEGF / EGF targeting antibody preferably contains a unit dosage form of VEGF / EGF targeting antibody, and the preparation containing chemotherapeutic drug preferably contains a unit dosage form of chemotherapeutic drug.

[0131] The step preferably places at least one unit dosage form containing recombinant human adenovirus type 5, at least one unit dosage form containing VEGF / EGF targeting antibody, and at least one unit dosage form containing chemotherapy drugs, as well as instructions, together to form a medicine box.

[0132] The main advantages of the present invention are:

[0133] (1) The combination of recombinant human adenovirus type 5 with VEGF / EGF targeting antibodies and chemotherapy drugs (such as FOLFOX6 / FOLFIRI) can synergistically and more efficiently inhibit colon cancer liver metastasis.

[0134] (2) After the drug combination of the present application was administered to patients with liver metastasis of colon cancer, no serious side effects occurred during and after the treatment, and no related dose-limiting toxicity (DLT) occurred, indicating good safety.

[0135] (3) Recombinant human adenovirus type 5 can stimulate a strong, systemic, and potentially long-lasting anti-tumor immune response, and can change the immune microenvironment of colorectal cancer liver metastases, which are relatively unresponsive to immunotherapy such as immune checkpoint inhibitors, from "cold" to "hot," making them effective for immunotherapy.

[0136] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the materials and reagents used in the embodiments of the present invention are all commercially available products. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions, such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989) The conditions described, or according to the conditions recommended by the manufacturer.

[0137] Unless otherwise indicated, percentages and parts are by weight.

[0138] Efficacy evaluation criteria for this application: mRECIST criteria

[0139]

[0140] Example 1

[0141] Clinical trial of recombinant human adenovirus type 5 combined with VEGF-targeted antibodies and chemotherapy drugs1

[0142] (1) Case introduction

[0143] Patient: Liu XX, male, 54 years old, underwent laparoscopic rectal DIXON"S surgery on May 7, 2018. Postoperative pathology showed: Radical specimen of rectal cancer: adenocarcinoma grade II, discoid type, tumor size 6.6×5.6×1.2cm, infiltrating the entire layer of the intestinal wall to the surrounding fat tissue; nerve invasion was seen, and no clear intravascular cancer thrombus was found; no cancer involvement was found at the resection margins at both ends of the specimen and the circumferential resection margin; no cancer metastasis was found in the highest group of lymph nodes (0 / 3) and lymph nodes around the intestinal wall (0 / 11); no tumor was found in the "anastomotic resection margin". After taking S-1 for one week after surgery, no other treatment was continued and regular follow-up was performed. The disease recurred in June 2019, and a "transanal resection of rectal tumor" was performed on June 20, 2019. Pathology suggested adenocarcinoma, and no adjuvant chemotherapy or other treatment was performed after the operation (details unknown). PET / CT was reviewed on September 26, 2019: 1. After "radical resection of rectal cancer", the FDG metabolism of the anastomosis increased, and the possibility of anastomotic inflammation was considered high; multiple metastases in the left upper lung field and the posterior segment of the right upper lobe, liver metastasis; left presacral soft tissue metastasis, bilateral inguinal lymph node metastasis. From December 11, 2019 to March 12, 2020, 5 cycles of chemotherapy with Erbitux + Capeox regimen: Erbitux 800mg d1+oxaliplatin 200mg d2+capecitabine 1.5bid orally d2-d14, efficacy evaluation PD. Pathology of right inguinal lymph node biopsy on March 10, 2020: adenocarcinoma metastasis with necrosis, considered intestinal origin based on medical history. RAS and BRAF gene testing indicated wild type.

[0144] Physical examination: clear consciousness, no icteric skin and sclera, flat and soft abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, edema of both lower limbs.

[0145] For further treatment, the patient was enrolled in the clinical study "Clinical study on the efficacy and safety of (Ancore) recombinant human adenovirus type 5 injection combined with standard treatment for liver metastasis of gastrointestinal malignancies" on April 7, 2020, excluding contraindications and signing an informed consent.

[0146] (2) Treatment options

[0147] On April 9, 2020, ultrasound-guided liver lesion puncture biopsy was performed. Pathology: (liver tumor puncture specimen) metastatic adenocarcinoma, combined with clinical history, intestinal origin was considered. Immunohistochemistry results: tumor cells ck (+), ck7 (+), ck20 (+), ck8 / 18 (+), ck19 (+), villin (+), cdx-2 (+), afp (-), he p-1 (-), ki67 (+75%).

[0148] On April 9 and 24, 2020, the right liver tumor was treated with oncolytic virus injection (1 ml of Ankorui), and on April 11, 2020, bevacizumab + FOLFIRI chemotherapy was performed: bevacizumab 400 mg d1 (5 mg / kg) + irinotecan 320 mg (180 mg / m 2 )d1+calcium folinate 0.7(400mg / m 2 )d1+ fluorouracil 0.7 (400 mg / m 2 )d1+ fluorouracil 4.0 (2400 mg / m 2 )civ46h,q2w.

[0149]

[0150] (3) Treatment effect

[0151] After the injection of Ankorui, the patient was in good condition and reported no obvious symptoms of discomfort.

[0152] The long diameter of the target lesion before and after treatment was 26.4 mm ( Figure 1 Left), 28.7mm( Figure 1 Right), imaging examination showed that the tumor increased by 8.7%, and the efficacy evaluation was SD.

[0153] Example 2

[0154] Clinical trial of recombinant human adenovirus type 5 combined with VEGF-targeted antibodies and chemotherapy drugs 2

[0155] (1) Case introduction

[0156] Patient Ding XX, male, 72 years old, had right upper abdominal pain in November 2019. Ultrasound was performed in an external hospital: multiple solid masses in the liver, and then upper abdominal CT was performed: 1. Multiple masses in the liver, considering the possibility of multiple metastases. 2. Irregular thickening of the intestinal wall in the proximal part of the ascending colon. Liver puncture pathology on December 9, 2019: poorly-moderately differentiated adenocarcinoma with large necrosis, gastrointestinal metastasis was considered first. C1-8 A+FOLFOX targeted chemotherapy was performed from December 12, 2019 to May 9, 2020: bevacizumab 300mg d0+oxaliplatin (Aiheng) 200mg d1+leucovorin 600mg d1+fluorouracil injection 3.5CIV 46h, 05-23 efficacy evaluation pd. During the period from May 29 to August 21, 2020, C1-6A+FOLFIRI targeted chemotherapy was given: bevacizumab 300mg d0+irinotecan 200mgd1+leucovorin 600mg d1+fluorouracil injection 3.5CIV 46h, and the efficacy was evaluated in August. During the same period from December 11, 2019 to June 30, 2020, liver HIFU treatment was given 5 times.

[0157] Physical examination: clear consciousness, no icteric skin and sclera, flat and soft abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, edema of both lower limbs.

[0158] For further treatment, the patient met the inclusion criteria of the project "Clinical Study on the Efficacy and Safety of (Ankorui) Recombinant Human Adenovirus Type 5 Injection Combined with Standard Treatment for the Treatment of Liver Metastasis of Gastrointestinal Malignancies". After communicating with the patient, the patient signed the informed consent form on September 1, 2020.

[0159] (2) Treatment options

[0160] Ultrasound-guided liver puncture biopsy was performed on September 1, 2020. Oncolytic virus injection (1 ml of Ankorui) was performed on September 1 and 17, 2020. C7-8 A+FOLFIRI targeted chemotherapy was given on September 3 and 19, 2020: bevacizumab 300 mg d1 + irinotecan 200 mg d1 + calcium folinate 300 mg d1 + fluorouracil injection 3.5 CIV 46 h.

[0161]

[0162]

[0163] (3) Treatment effect

[0164] After the second injection of Ankorui, the patient developed fever, with the highest temperature reaching 39.2℃. Physical cooling and indomethacin suppositories were given to treat the symptoms. The patient's temperature subsequently dropped and he had no other discomfort.

[0165] The long diameter of the target lesion before and after treatment was 28.3 mm ( Figure 2 Left), 27.2mm( Figure 2 Right), imaging examination showed that the tumor was reduced by 3.9%, and the efficacy evaluation was SD.

[0166] Example 3

[0167] Clinical trial of recombinant human adenovirus type 5 combined with VEGF-targeted antibodies and chemotherapy drugs

[0168] (1) Case introduction

[0169] Patient Zhu XX, male, 64 years old, had a change in bowel habits and mucous bloody stools for more than half a year at the end of 2012. On December 12, he underwent "laparoscopic sigmoid colon tumor radical resection + laparoscopic intestinal adhesion lysis". [Postoperative pathology] (sigmoid colon) moderately differentiated tubular adenocarcinoma, partial mucinous adenocarcinoma, invading the entire layer of the intestinal wall, no tumor involvement was found in the upper and lower cut ends, (lymph nodes adjacent to the mass) lymph node metastatic adenocarcinoma (3 / 16), some tumors invaded the lymph node capsule, and local cancer nodules were formed. Postoperative Xelox chemotherapy was performed 5 times (dose unknown). Liver metastasis was found during follow-up on December 18, 2017, and no further diagnosis and treatment was given. On November 20, 2020, a liver mass puncture biopsy was performed at the Oriental Hepatobiliary Hospital. [Pathology] (liver biopsy) adenocarcinoma, moderately differentiated, considered to be from the intestine. Genetic testing: KRAS / NRAS gene mutation combined detection: KRAS Exon-2 G12C, G12R, G12V, G12A, G13C mutations, the remaining sites detected as wild type; BRAF gene mutation detection Exon-15 V600E wild type; MSI: MSS. PET / CT examination at the same time considered liver metastasis. From December 28, 2020, bevacizumab + FOLFIRI targeted chemotherapy was given: bevacizumab 300mg (7.5mg / kg) d0 + irinotecan 280mg (180mg / m2) d1 + leucovorin 700mg (400mg / m2) d1 + fluorouracil 700mg (400mg / m2) d1 + fluorouracil 3.5 (2.4 / m2) civ 46 hours.

[0170] Physical examination: clear consciousness, no icteric skin and sclera, flat and soft abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, edema of both lower limbs.

[0171] Because the patient met the inclusion criteria of the project "Clinical Study on the Efficacy and Safety of (Ancore) Recombinant Human Adenovirus Type 5 Injection Combined with Standard Treatment for Liver Metastasis of Gastrointestinal Malignancies", after communicating with the patient, the patient signed the informed consent on January 8, 2021.

[0172] (2) Treatment options

[0173] Liver puncture biopsy was performed on January 8, 2021. Oncolytic virus injection therapy was performed on January 8, 29, 19, and 19, 2021. During the same period, bevacizumab + FOLFIRI (C2-4) was given on January 12, 1, 22, and 22, 2021: bevacizumab 300 mg (7.5 mg / kg) d0 + irinotecan 280 mg (180 mg / m2) d1 + folinic acid 700 mg (400 mg / m2) d1 + fluorouracil 700 mg (400 mg / m2) d1 + fluorouracil 3.5 (2.4 / m2) civ 46 hours of targeted chemotherapy.

[0174]

[0175] (3) Treatment effect

[0176] After the first and third injections of Ankorui, the patient developed fever, with the highest temperature being 38.5°C. After antipyretic treatment, the temperature dropped and there was no other discomfort.

[0177] The long diameter of the target lesion before and after treatment was 30.8 mm ( Figure 3 Left), 28.6m( Figure 3 Right), imaging examination showed that the tumor was reduced by 7.14%, and the efficacy evaluation was SD.

[0178] Example 4

[0179] Clinical trial of recombinant human adenovirus type 5 combined with VEGF-targeted antibodies and chemotherapy drugs

[0180] (1) Case introduction

[0181] Patient Zhang, male, 63 years old, visited a local hospital for abdominal pain in December 2019. PET-CT showed: ascending colon cancer with mesenteric and presacral lymph node metastasis, and multiple liver metastases. From January 2, 2020 to May 18, 2020, he received 6 cycles of Avastin + XELOX chemotherapy. From June 18, 2020 to August 20, 2020, he was treated with FOLFOX + Erbitux. Later, due to the inability to undergo surgery, he continued to receive Avastin + XELOX chemotherapy from September 21, 2020 to May 19, 2021, and is now hospitalized for continued treatment.

[0182] Physical examination: Observation: The whole abdomen is flat, the abdominal wall veins are not exposed, abdominal breathing can be felt, no gastrointestinal peristaltic waves are seen, and no intestinal pattern is felt. Palpation: The abdominal muscles are soft, no tenderness, no rebound pain, the liver and spleen are not palpable under the ribs, Murphys' (-). Percussion: No shifting dullness, no percussion pain in the kidney area, no percussion pain in the liver area. Auscultation: Normal bowel sounds, 4 times / min.

[0183] To further reduce the tumor, patients are recommended to join the "Clinical study on the effectiveness and safety of (Ankorui) recombinant human adenovirus type 5 injection combined with standard treatment for liver metastasis of gastrointestinal malignancies" project.

[0184] (2) Treatment options

[0185] The patient was injected with 1 ml of Ankorui into the liver lesion under ultrasound guidance on June 21, 2021 and July 26, 2021, for a total of two cycles. The day after the injection of Ankorui, the treatment regimen of Avastin + Capecitabine (bevacizumab 540 mg + capecitabine 1.5 g orally bid d1-14) was carried out.

[0186]

[0187] (3) Treatment effect

[0188] After the injection of Ankorui, the patient was in good condition and reported no obvious symptoms of discomfort.

[0189] The target lesion volume was reduced from 15.6 mm to 11.4 mm before and after treatment. Figure 4 ), imaging examination showed that the tumor was significantly reduced, and the efficacy was evaluated as PR (partial remission).

[0190] Example 5

[0191] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs1

[0192] (1) Case introduction

[0193] Patient Zhang X, female, 50 years old, had a change in bowel habits since October 2019. Colonoscopy was performed to consider colonic mass, but the patient did not pay attention to it later. Colonoscopy was performed again in the hospital on April 28, 2020. Pathology: rectal adenocarcinoma. Genetic testing: KRAS / NRAS, BRAF gene mutation (-). Imaging examinations considered liver, lung, and lymph node metastasis of rectal malignant tumors. From May 9 to August 31, 2020, chemotherapy with C1-6 Erbitux + SOX regimen was performed: Cetuximab 700mg (500mg / m2) d1 + Oxaliplatin 100-175mg (85mg / m2) d1 + S-1 50mg bid d1-d14, q3w. During the period, the efficacy evaluation was PR on July 2020, and SD on September. During the period from 2020-05-07 to 09-02, HIFU treatments for liver lesions were performed 6 times.

[0194] Physical examination: clear consciousness, no icteric skin and sclera, flat and soft abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, edema of both lower limbs.

[0195] Because the patient met the inclusion criteria of the project "Clinical Study on the Efficacy and Safety of (Ancore) Recombinant Human Adenovirus Type 5 Injection Combined with Standard Treatment for Liver Metastasis of Gastrointestinal Malignancies", after communicating with the patient, the patient signed an informed consent form on September 23, 2020 to be included in the group.

[0196] (2) Treatment options

[0197] Ultrasound-guided liver puncture biopsy was performed on September 23, 2020. Ultrasound-guided oncolytic virus immunotherapy (1 ml of Ankerui) was performed on September 23 and October 28, 2020. C7-8 Erbitux + SOX chemotherapy was performed on September 26 and October 30, 2020: Cetuximab 700 mg (500 mg / m 2 )d1+oxaliplatin 175mg (130mg / m 2 )d1+ S-1 50mg bid d1-d14, q3w;

[0198]

[0199] In addition, highly focused ultrasound (HIFU) treatment of liver lesions was performed on 2020-09-23 and 2020-10-28.

[0200] (3) Treatment effect

[0201] After the injection of Ankorui, the patient was in good condition and reported no obvious symptoms of discomfort.

[0202] The long diameter of the target lesion before and after treatment was 29.4 mm ( Figure 5 Left), 26.9mm( Figure 5 Right), imaging examination showed that the tumor was reduced by 8.5%, and the efficacy evaluation was SD.

[0203] Example 6

[0204] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs 2

[0205] (1) Case introduction

[0206] Patient Chen XX, male, 61 years old, developed abdominal pain on May 2, 2020. Ultrasound was performed to consider intestinal obstruction and intrahepatic mass: he was treated on May 8, 2020, and imaging examinations considered multiple liver mass, colon cancer, and nodules in both lungs. On May 14, 2020, he underwent radical resection of the hepatic flexure tumor of the colon under general anesthesia. Postoperative pathology: (transverse colon) adenocarcinoma, low-to-moderate differentiation, infiltrative ulcerative type, cancer infiltrates the entire layer of the intestinal wall to the serosa, and vascular cancer thrombus and nerve invasion can be seen; no cancer involvement was found in the upper and lower resection margins of the specimen and the omental tissue. Lymph nodes (peri-intestinal lymph nodes) were found to be involved in cancer (6 / 8), and 13 cancer nodules were found. Genetic testing: KRAS / NRAS, BRAF gene wild type, MSI: MSS. From June 28 to July 28, 2020, C1-3 cetuximab + mFOLFOX6 targeted chemotherapy: Erbitux 800 mg (500 mg / m 2 )d0+oxaliplatin 125mg (85mg / m 2 )d1+calcium folinate 600mg(400mg / m 2 )d1+ fluorouracil 600mg (400mg / m 2 )d1+ fluorouracil 3.5 (2400 mg / m 2 ) The patient developed a rash after receiving chemotherapy with the Civ 46h q2w regimen. From August 11, 2020 to November 21, 2020, he received C4-10 cetuximab (reduced dose) + mFOLFOX6 targeted chemotherapy: Erbitux 400mg (250mg / m 2 )d0+oxaliplatin 125mg (85mg / m 2 )d1+calcium folinate 600mg(400mg / m 2 )d1+ fluorouracil 600mg (400mg / m 2 )d1+ fluorouracil 3.5 (2400 mg / m 2 )civ 46h q2w chemotherapy. The efficacy evaluation was reviewed in October 2020 and the result was SD.

[0207] Physical examination: clear consciousness, no icteric skin and sclera, flat and soft abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, edema of both lower limbs.

[0208] Because the patient met the inclusion criteria of the project "Clinical Study on the Efficacy and Safety of (Ancore) Recombinant Human Adenovirus Type 5 Injection Combined with Standard Treatment for Liver Metastasis of Gastrointestinal Malignancies", after communicating with the patient, the patient signed the informed consent on November 4, 2020.

[0209] (2) Treatment options

[0210] On November 4 and 19, 2020, he received oncolytic virus injection therapy for liver tumors (2 ml of Ankerui for the first time and 4 ml for the second time). On November 5 and 21, 2020, he received C9-10 cetuximab (reduced dose) + mFOLFOX6 targeted chemotherapy: Erbitux 400 mg (250 mg / m 2 )d0+oxaliplatin 125mg (85mg / m 2 )d1+calcium folinate 600mg(400mg / m 2 )d1+ fluorouracil 600mg (400mg / m 2 )d1+ fluorouracil 3.5 (2400 mg / m 2 )civ 46h q2w chemotherapy.

[0211]

[0212]

[0213] (3) Treatment effect

[0214] After the second injection of Ankorui, the patient developed fever, with the highest temperature being 39.7℃. After antipyretic treatment, the temperature dropped and there was no other discomfort.

[0215] The long diameter of the target lesion before and after treatment was 28.3 mm ( Figure 6 Left), 27.2mm( Figure 6 Right), imaging examination showed that the tumor was reduced by 3.9%, and the efficacy evaluation was SD.

[0216] Example 7

[0217] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs

[0218] (1) Case introduction

[0219] Patient Zhou XX, female, 71 years old, was diagnosed with "edema of both lower limbs and difficulty walking" before March 2021. Imaging examinations showed multiple lesions in the liver, and metastatic tumors were considered. [Liver puncture] (2021.03.15) (liver puncture specimen) metastatic adenocarcinoma, he morphology combined with immunohistochemistry results first considered gastrointestinal origin, please combine clinical and imaging. 2021-3-12 colonoscopy showed descending colon tumor, [pathology] (colon) high-grade intraepithelial neoplasia in some glandular epithelial cells, local carcinoma. Because pathological gene testing could not be performed due to tissue sampling, and the patient had poor tolerance, CapOX was performed on 2021-3-19: oxaliplatin 150mg (130mg / m2) d1 + capecitabine 1.0g bid d1-14 q3w chemotherapy. No mutations were found in peripheral blood RAS and RAF. On April 9, 2021 and April 28, 2021, cetuximab + CapOX: cetuximab 700 mg (500 mg / m2) d1 + oxaliplatin 150 mg (130 mg / m2) d1 + capecitabine 1.0 g bid d1-14 q3w. The efficacy was reviewed in May 2021 and the result was PR. The patient's general condition improved, and the plan was changed to C1-2 cetuximab 700mg (500mg / m2) d1 + oxaliplatin 150mg (135mg / m2) d1 + calcium folinate 500mg (400mg / m2) d1 + fluorouracil 500mg (400mg / m2) d1 + fluorouracil 3.0 (2400mg / m2) civ 46hq2w, targeted chemotherapy on 2021-05-20 and 06-02.

[0220] For further treatment, the patient was enrolled in the "Clinical Study on the Efficacy and Safety of (Ankorui) Recombinant Human Adenovirus Type 5 Injection Combined with Standard Treatment for Liver Metastasis of Gastrointestinal Malignancies". After communicating with the patient, the patient signed the informed consent on June 16, 2021.

[0221] (2) Treatment options

[0222] On June 16, 2021, a liver biopsy was performed. On June 16 and July 2, 2021, oncolytic virus (1 ml of Ankorui) was injected into the liver lesions. On June 18 and July 4, 2021, C3-4 cetuximab 700 mg (500 mg / m 2 )d1+oxaliplatin 150mg (135mg / m 2 )d1+calcium folinate 500mg(400mg / m 2 )d1+500mg fluorouracil (400mg / m 2 )d1+ fluorouracil 3.0 (2400 mg / m 2 )civ 46h q2w, targeted chemotherapy.

[0223]

[0224] (3) Treatment effect

[0225] The patient had a fever of 38.4 degrees on the day of the first injection of Ankori, and his condition improved after taking Tylenol.

[0226] After the last chemotherapy, the patient developed grade IV bone marrow suppression, which improved with remdesivir treatment.

[0227] After the second injection of Ankorui, the patient developed fever, with the highest temperature being 38.9℃. After antipyretic treatment, the temperature dropped and there was no other discomfort.

[0228] The long diameter of the target lesion before and after treatment was 46.5 mm ( Figure 7 Left), 36.6m( Figure 7 Right), imaging examination showed that the tumor was reduced by 21.3%, and the efficacy evaluation was SD.

[0229] Example 8

[0230] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs

[0231] (1) Case introduction

[0232] Patient Xu, male, 67 years old, had colonoscopy in our hospital on April 15, 2021 due to changes in bowel habits: multiple polyps in the colorectum. Pathology suggests: malignant tumor of the ascending colon. Further abdominal enhanced CT and liver MRI examinations showed: multiple liver metastases. On May 7, 2021 and May 27, 2021, he underwent chemotherapy with Erbitux + FOLFOX regimen, and is now admitted to the hospital for the next cycle of chemotherapy.

[0233] Physical examination: clear consciousness, good spirit, flat abdomen, no exposed abdominal wall veins, abdominal breathing, no gastrointestinal peristaltic waves, no intestinal type. Palpation: soft abdominal muscles, no tenderness, no rebound pain, liver and spleen are not palpable under the ribs, Murphys' (-). Percussion: no mobile dullness, no percussion pain in the kidney area, no percussion pain in the liver area. Auscultation: normal bowel sounds, 4 times / min.

[0234] To further reduce the tumor, patients are recommended to join the "Clinical study on the effectiveness and safety of (Ankorui) recombinant human adenovirus type 5 injection combined with standard treatment for liver metastasis of gastrointestinal malignancies" project.

[0235] (2) Treatment options

[0236] The patient was injected with 2 ml of Ankorui into the liver lesion under ultrasound guidance on June 18, 2021 and July 16, 2021, for a total of two cycles. The next day after the injection of Ankorui, the chemotherapy regimen of Erbitux (1000 mg) + FOLFOX (oxaliplatin 150 mg intravenous drip d1, calcium folinate 400 mg intravenous drip d1, fluorouracil 500 mg intravenous push d1, fluorouracil 3500 mg maintenance 48 hours) was continued.

[0237]

[0238] (3) Treatment effect

[0239] After the injection of Ankorui, the patient was in good condition. He developed a fever that night with a body temperature of 37.8℃. He did not receive symptomatic treatment and recovered on his own without any other discomfort symptoms.

[0240] The target lesion volume was reduced from 47.7 mm to 43.3 mm before and after treatment. Figure 8 ), imaging examination showed that the tumor was reduced, and the efficacy was evaluated as SD (stable disease).

[0241] Example 9

[0242] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs

[0243] (1) Case introduction

[0244] Patient Xu, male, 59 years old, went to a local hospital for treatment due to abdominal discomfort. PET-CT showed: sigmoid colon cancer with multiple lymph node metastases and liver metastases. He received FOLFOXIRI chemotherapy on April 14, 2021. From April 28, 2021 to June 12, 2021, he received FOLFOXIRI+Erbitux treatment. He is now hospitalized for continued treatment.

[0245] Physical examination: clear consciousness, good spirit, flat abdomen, no exposed abdominal wall veins, abdominal breathing, no gastrointestinal peristaltic waves, no intestinal type. Palpation: soft abdominal muscles, no tenderness, no rebound pain, liver and spleen are not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area, no percussion pain in the liver area. Auscultation: normal bowel sounds, 4 times / min.

[0246] To further reduce the tumor, patients are recommended to join the "Clinical study on the effectiveness and safety of (Ankorui) recombinant human adenovirus type 5 injection combined with standard treatment for liver metastasis of gastrointestinal malignancies" project.

[0247] (2) Treatment options

[0248] The patient was injected with 1 ml of Ankori into the liver lesion under ultrasound guidance on June 29, 2021 and July 20, 2021, for a total of two cycles. On the second day after Ankori injection, the FOLFOXIRI (irinotecan 200 mg intravenous drip d1, oxaliplatin 140 mg intravenous drip d1, calcium folinate 400 mg intravenous drip d1, fluorouracil 3500 mg for 48 hours) + Erbitux (800 mg) treatment regimen was performed.

[0249]

[0250] (3) Treatment effect

[0251] After the injection of Ankorui, the patient was in good condition and reported no obvious symptoms of discomfort.

[0252] Before and after treatment, the target lesion volume was reduced from 38.0 mm to 23.9 mm. Imaging examinations showed that the tumor was significantly reduced, and the efficacy was evaluated as PR (partial remission).

[0253] Example 10

[0254] Clinical trial of recombinant human adenovirus type 5 combined with EGF-targeted antibodies and chemotherapy drugs6

[0255] (1) Case introduction

[0256] Patient Sun, male, 56 years old. On September 11, 2020, he went to the local hospital for colonoscopy, which showed moderately differentiated adenocarcinoma. PET-CT showed colon cancer, multiple low-density metastatic lesions in the liver. On September 12, 2020, he underwent chemotherapy with FOLFOX+bevacizumab. From September 27, 2020 to June 13, 2021, he went to Changhai Hospital for 13 cycles of chemotherapy with FOLFOX+cetuximab. He is now admitted to the hospital for further treatment.

[0257] Physical examination: clear consciousness, no yellowing of skin and mucous membranes. Flat abdomen, no tenderness, no rebound pain, liver and spleen not palpable under the ribs, Murphys' (-). Percussion: no shifting dullness, no percussion pain in the kidney area. No abnormal bowel sounds, 4 times / min.

[0258] Because the patient met the inclusion criteria of the project "Clinical study on the effectiveness and safety of (Ancore) recombinant human adenovirus type 5 injection combined with standard treatment for liver metastasis of gastrointestinal malignancies", after communication with the patient, the patient signed an informed consent form for inclusion in the group.

[0259] (2) Treatment options

[0260] The patient was injected with 1 ml of Ankorui into the tumor under ultrasound guidance on June 10, 2021, for a total of one cycle. After the injection of Ankorui, the patient continued with FOLFOX+cetuximab chemotherapy.

[0261]

[0262] (3) Treatment effect

[0263] After the injection of Ankorui, the patient developed fever.

[0264] The volume of the target lesion was reduced from 21.5 mm to 17.6 mm before and after treatment. Imaging examinations showed ablation of lesions around the target lesion, and the efficacy was evaluated as SD (stable disease).

[0265] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. Use of an active ingredient combination, characterized in that: The active ingredient combination includes: The first active ingredient is oncolytic virus, the second active ingredient is VEGF / EGF targeting antibody and the third active ingredient is chemotherapeutic drug, and the combination is used to prepare a pharmaceutical composition or a drug kit for synergistically treating liver metastasis of colon cancer.

2. The use according to claim 1, characterized in that The oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

3. The use according to claim 1, characterized in that The VEGF / EGF targeting antibody is selected from the group consisting of bevacizumab, cetuximab, or a combination thereof.

4. The use according to claim 1, characterized in that The chemotherapy drug is selected from the following group: irinotecan, oxaliplatin, folinic acid, fluorouracil, capecitabine, S-1, or a combination thereof.

5. The use according to claim 1, characterized in that The oncolytic virus was injected at a rate of 0.5×10 12 VP to 2.5×10 12 VP is administered at a dose of 1 to 20 mg / kg, preferably 2 to 15 mg / kg, and more preferably 3 to 10 mg / kg.

6. An active ingredient combination, characterized in that The combination consists of an oncolytic virus, a VEGF / EGF targeting antibody and a chemotherapy drug.

7. The active ingredient combination according to claim 6, characterized in that The oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

8. The active ingredient combination according to claim 6, characterized in that The VEGF / EGF targeting antibody is selected from the group consisting of bevacizumab, cetuximab, or a combination thereof.

9. A pharmaceutical composition for treating liver metastasis of colon cancer, characterized in that: The pharmaceutical composition comprises (a) oncolytic virus, (b) VEGF / EGF targeting antibody, (c) chemotherapeutic drug and (d) pharmaceutically acceptable carrier.

10. A medicine box, comprising: (a) a first preparation, wherein the first preparation contains an oncolytic virus and a pharmaceutically acceptable carrier; (b) a second formulation comprising a VEGF / EGF targeting antibody and a pharmaceutically acceptable carrier; (c) a third preparation comprising a chemotherapeutic drug and a pharmaceutically acceptable carrier; and (d) instructions describing a method for treating colon cancer liver metastasis by combining an oncolytic virus, a VEGF / EGF targeting antibody, and a chemotherapeutic drug.

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