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EP-4737563-A1 - RECOMBINANT ONCOLYTIC VACCINIA VIRUS AND USE THEREOF

EP4737563A1EP 4737563 A1EP4737563 A1EP 4737563A1EP-4737563-A1

Abstract

Provided in the present invention is a recombinant oncolytic vaccinia virus, which is operably inserted into a synonymously mutated exogenous gene capable of expressing 4-1BBL, and also provided is the use of the recombinant oncolytic vaccinia virus in the preparation of a drug for preventing or treating tumors and cancers. The present invention has the following beneficial effects: the synonymous-mutation-based recombinant vaccinia virus VV-mH4-1BBL retains the original oncolytic effect of the oncolytic virus and the functions thereof of initiating and enhancing anti-tumor immune responses, and improves the safety by means of deleting the TK gene; 4-1BBL is highly expressed on the surface of a tumor cell, such that 4-1BBL can enhance anti-tumor immunity by means of exciting a 4-1BB signal of 4-1BB + immune cells (including T cells) in the tumor microenvironment, and 4-1BBL is also confined within tumor tissues to exert the function thereof in a centralized manner, thereby avoiding potential systemic toxic side effects; and the introduction of a synonymous mutation site enables the virus to detect the expression of a therapeutic (exogenous) 4-1BBL gene during treatment.

Inventors

  • JU, Songguang
  • GE, Yan

Assignees

  • Suzhou Onlyv Biotechnology Limited Company

Dates

Publication Date
20260506
Application Date
20231220

Claims (10)

  1. A recombinant oncolytic vaccinia virus, characterized in that the recombinant oncolytic vaccinia virus is operably inserted into a synonymously mutated exogenous gene capable of expressing 4-1BBL.
  2. The recombinant oncolytic vaccinia virus according to claim 1, characterized in that the exogenous gene is inserted into TK gene of the vaccinia virus.
  3. The recombinant oncolytic vaccinia virus of claim 1, characterized in that DNA sequence of the exogenous gene is shown as SEQ ID No.1 to SEQ ID No.3.
  4. The recombinant oncolytic vaccinia virus according to claim 1, characterized in that amino acid sequence of the exogenous gene is shown as SEQ ID No.4.
  5. The recombinant oncolytic vaccinia virus according to claim 1, characterized in that the exogenous gene is linked via an IRES sequence.
  6. The recombinant oncolytic vaccinia virus according to claim 5, characterized in that the IRES sequence is shown as SEQ ID No.5.
  7. A pharmaceutical composition, characterized in that the pharmaceutical composition comprises the recombinant oncolytic vaccinia virus according to any one of claims 1-6, a pharmaceutically acceptable carrier and a pharmaceutical excipient.
  8. The pharmaceutical composition according to claim 7, characterized in that the administration mode of the pharmaceutical composition is direct injection and/or injection in combination with an endoscope; and the endoscope is preferably a laparoscope, a choledochoscope, a thoracoscope, an enteroscope, and a neuroendoscope.
  9. Use of the recombinant oncolytic vaccinia virus according to any one of claims 1-6 or the pharmaceutical composition according to any one of claims 7-8 in the preparation of a drug for preventing or treating a tumor and/or a cancer.
  10. The use according to claim 9, characterized in that the tumor and/or cancer is a solid tumor, preferably a cold tumor and/or a hot tumor, and the histological types thereof include, but are not limited to, pancreatic cancer, gallbladder cancer, liver cancer, colorectal cancer, gastric cancer, esophageal cancer, brain glioma, ovarian cancer, cervical cancer, prostate cancer, renal cancer, lung cancer, breast cancer, multiple myeloma, lymphoma, and melanoma.

Description

Technical Field The invention relates to the technical field of biomedicine, in particular to a recombinant oncolytic vaccinia virus and use thereof. Background Stimulation of 4-1BB signaling in immune cells can elicit strong and sustained anti-tumor immunity. At present, 4-1BB agonistic monoclonal antibodies, such as urelumab and utomilumab, have entered clinical research. When these antibodies are used systemically, because of the small difference between the effective dose and the safe dose, systemic toxic side effects often occur when the anti-tumor immune effect is effectively stimulated, or the anti-tumor effect is not significant when there are no obvious toxic side effects. These contradictions greatly limit their clinical application. Soluble 4-1BBL can stimulate 4-1BB signal, but is not used in clinical application studies. If it is directly used systemically, it can strongly enhance the immune response in the whole body, resulting in serious toxic side effects. Oncolytic viruses can selectively infect tumor cells, induce immunogenic death of tumor cells, initiate anti-tumor immunity and cause local inflammation, and act as immune adjuvants. However, oncolytic virus alone is often used to treat tumors because the antiviral effect of the body can quickly eliminate the virus, so it is necessary to enhance the anti-tumor immunity triggered by oncolytic virus after it plays its role in order to induce strong and sustained anti-tumor effects. Therefore, it is necessary to introduce immune enhancing factors to improve the anti-tumor effect of oncolytic viruses. It has been reported that mouse 4-1BBL recombinant oncolytic virus was constructed by using mouse 4-1BBL gene, and its reliability was proved by mouse model. However, recent studies have found that the wild-type 4-1BBL gene is commonly expressed in tumor cells such as intestinal cancer, and its encoding product (4-1BBL molecule) is located in the cells but not on the surface the cells. Therefore, if the wild-type 4-1BBL gene is used to construct a 4-1BBL recombinant oncolytic virus for treatment, the expression and localization of therapeutic 4-1BBL will not be detected and tracked. Oncolytic virus can selectively infect and cause immunogenic death of tumor cells, and then trigger the body's anti-tumor immune response, making it a feasible means of tumor immunotherapy, thereby to enter the field of tumor immune drug research and development. However, oncolytic virus itself is easy to be cleared by the body, and the anti-tumor immunity triggered by oncolytic virus is facing the common problem of being blocked by negative mechanisms such as immune checkpoints. Therefore, the overall design idea in the field is to construct recombinant oncolytic viruses, retain the advantages of the original oncolytic virus, avoid the weaknesses thereof, and then introduce immune enhancing factors to overcome the negative immune mechanism and induce safe, effective, systemic and sustained anti-tumor immunity. Costimulatory molecule ligand 4-1BBL strongly promotes the body's anti-tumor immunity by acting on 4-1BB molecules on the surface of T cells, NK cells, DC and so on, that is, by 4-1BB/4-1BBL pathway. Therefore, the biological agents taking 4-1BB/4-1BBL pathway as a target may become a new type of anti-tumor drugs. Agonistic 4-1BB monoclonal antibody and 4-1BBL recombinant oncolytic virus are important biological agents in the field, but their shortcomings restrict their future application and need to be improved. The agonistic 4-1BB monoclonal antibody can activate the 4-1BB/4-1BBL pathway to produce strong anti-tumor effect, but the dosage difference between the effective anti-tumor effect and the toxic side effect of the agonistic 4-1BB monoclonal antibody is small, that is, the safe dosage range is small. Thus, the selection of the natural ligand 4-1BBL as an immune activator of the 4-1BB molecule provides a natural stimulus to activate the 4-1BB signal. In addition, if soluble 4-1BBL is directly used to treat tumors, it may bring toxic side effects due to the systemic effects of cytokines. However, it has been reported that 4-1BBL molecules generally exist in intestinal cancer, pancreatic cancer and other tumor cells, but are not expressed in the cell membrane, so that tumor cells can avoid stimulating 4-1BB molecules on the surface of immune cells, which is a mechanism of tumor immune escape. If the natural ligand 4-1BBL is used to construct a recombinant oncolytic virus, 4-1BBL can be expressed on the surface of tumor cells to stimulate anti-tumor immunity. However, in the future treatment process, it is impossible to distinguish whether the 4-1BBL gene in the tumor tissue comes from the tumor tissue itself or the therapeutic 4-1BBL recombinant oncolytic virus, which makes it extremely difficult to monitor and evaluate the efficacy. Summary of the Invention Aiming at the existing technical limitation as mentioned above, the invention provides a recombinan