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Intravenous oncolytic virus preparation and preparation method thereof

A technology of oncolytic virus and limited dilution method, which is applied in the direction of resisting vector-borne diseases, medical preparations containing active ingredients, and medical raw materials derived from viruses/bacteriophages, etc., can solve problems such as obstacles to the development of cancer virus therapy, and achieve good results. Universal applicability, improved therapeutic effect, and low immunogenicity

Active Publication Date: 2019-02-01
厦门宏谱福生物科技有限公司
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Problems solved by technology

However, when the oncolytic virus directly enters the body's blood circulation system, it will cause the body's inherent antiviral immunity and innate immune response, and oncolytic viruses are prone to non-specific isolation in tissues and organs, making traditional oncolytic virus vaccines impossible. Entering the body by intravenous injection poses a major hurdle to the development of cancer viral therapies

Method used

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  • Intravenous oncolytic virus preparation and preparation method thereof
  • Intravenous oncolytic virus preparation and preparation method thereof
  • Intravenous oncolytic virus preparation and preparation method thereof

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preparation example

[0034] Preparation example: the reagents used in the following synthetic steps are commercially available.

[0035] A method for preparing an oncolytic adenovirus vaccine wrapped in cell membrane vesicles modified by targeting polypeptide hepatitis B virus capsid pre-S1 protein (preS1), comprising the following steps:

[0036] 1. Preparation of cell membrane vesicles modified by targeting polypeptide PreS1:

[0037] 1) 24 hours before transfection, HepG2 cells in the logarithmic growth phase were digested with trypsin, and the cell density was adjusted to 5×10 per ml with a medium containing 10% serum. 5 Cells were re-seeded in a 10cm cell culture dish at 37°C, 5% CO 2 Cultured in an incubator. When the cell density reaches 70%-80%, it can be used for transfection, and the cell culture medium is replaced with serum-free medium before transfection;

[0038] 2) Mix the diluted eukaryotic expression plasmid containing the PreS1 gene with the commercial transfection reagent Lip...

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Abstract

The invention relates to an intravenous oncolytic virus preparation and a preparation method thereof. The method performs transfection of a targeting polypeptide gene to a eukaryotic cell, and displays the targeting polypeptide on a cell membrane to obtain the gene engineering reconstructed cell strain capable of stably expressing the targeting polypeptide, The cell strain is used to extract nano-sized cell membrane vesicles with a uniform size, and then the oncolytic virus (OV) is loaded into the cell membrane vesicles with engineering reconstruction to synthesize an intravenous oncolytic virus complex which can be targeted for tumor delivery. The intravenous oncolytic virus preparation encapsulates the oncolytic virus in the cell membrane vesicles, and can effectively reduce the neutralizing effect of a neutralizing antibody on the oncolytic virus in the body, the cell membrane vesicle has targeting performance after gene engineering reconstruction, can achieve targeted delivery of the oncolytic virus and enrichment at the tumor site, and reduces the systemic side effects of the oncolytic virus.

Description

technical field [0001] The invention relates to a preparation method of an intravenously injectable oncolytic virus vaccine. Background technique [0002] Oncolytic virus therapy is an ideal treatment for cancer. Taking advantage of the ability of viruses to selectively replicate and amplify tumor cells, oncolytic virus vaccines have achieved good therapeutic effects in a number of preclinical and clinical trials. Recently, the US FDA approved Amgen's oncolytic virus vaccine (T-VEC) for the treatment of skin and lymph node melanoma lesions. In addition, the use of oncolytic viruses in localized cancer gene therapy has been reported in several human clinical trial reports. However, when the oncolytic virus directly enters the body's blood circulation system, it will cause the body's inherent antiviral immunity and innate immune response, and oncolytic viruses are prone to non-specific isolation in tissues and organs, making traditional oncolytic virus vaccines impossible. E...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/76A61K35/761A61K9/08A61P35/00
CPCA61K9/0019A61K9/08A61K35/76A61K35/761A61P35/00Y02A50/30
Inventor 刘刚吕鹏刘旋陈笑梅刘超牟雅琳
Owner 厦门宏谱福生物科技有限公司
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