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Seneca valley virus based compositions and methods for treating disease

A virus genome, virus technology, applied in the direction of medical materials derived from viruses/phages, biochemical equipment and methods, viruses, etc.

Active Publication Date: 2006-12-06
NOVARTIS PHARM AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Despite the early promise of these newly defined classes of anticancer treatments, several limitations limit their utility as cancer treatments

Method used

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  • Seneca valley virus based compositions and methods for treating disease
  • Seneca valley virus based compositions and methods for treating disease
  • Seneca valley virus based compositions and methods for treating disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0243] The following examples are given to illustrate the present invention, and the present invention is not limited to the examples.

example 1

[0245] Amplification and purification of virus

[0246] To incubate SVV in PER.C6 cells:

[0247] SVV was plaque purified once and well-separated plaques were selected and expanded in PER.C6 cells (Fallaux et al., 1998). Crude virus lysate (CVL) from SVV-infected PER.C6 cells can be generated by 3 cycles of freezing and thawing and used to infect PER.C6 cells. PER.C6 cells were treated with Dulbecco's modified Eagle medium (DMEM, Invitrogen, Carlsbad, CA, USA) in 50×150cm 2 Incubation was carried out in a T.C flask. Infected cells were harvested 30 hours after infection when complete CPE was observed and collected by centrifugation at 1500 rpm, 4°C for 10 minutes. Cell pellets were resuspended in cell culture supernatant (30 ml) and subjected to 3 cycles of freezing and thawing. The resulting CVL was clarified by centrifugation at 1500 rpm for 10 minutes at 4°C. SVV was purified by 2 rounds of CsCl gradients: one step gradient centrifugation (CsCl density 1.24 g / ml and 1...

example 2

[0249] electron microscopy

[0250] SVVs were mounted on polymethylvinyl acetate carbon-coated grids using the direct application method, stained with uranyl acetate and examined by transmission electron microscopy. Representative photomicrographs of viruses were taken at high magnification. For transmission electron microscopy, ultrathin sections of SVV-infected PER.C6 cells were cut from embedding blocks and the resulting sections were examined by transmission electron microscopy.

[0251] Purified SVV particles are spherical and approximately 27 nm in diameter, and appear singly or in small aggregates on the grid. Representative images of SVV are shown in figure 2 middle. In some places, ruptured virions and empty capsids, which are permeable to the dye, are also visible. Ultrastructural studies of infected PER.C6 cells revealed crystalline inclusions in the cytoplasm. Representative images of SVV-infected PER.C6 cells are shown in image 3 middle. Virus-infected ce...

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Abstract

The present invention relates to a novel RNA picornavirus that is called Seneca Valley virus ('SVV'). The invention provides isolated SVV nucleic acids and proteins encoded by these nucleic acids. Further, the invention provides antibodies that are raised against the SVV proteins. Because SVV has the ability to selectively kill some types of tumors, the invention provides methods of using SVV and SVV polypeptides to treat cancer. Because SVV specifically targets certain tumors, the invention provides methods of using SVV nucleic acids and proteins to detect cancer. Additionally, due to the information provided by the tumor-specific mechanisms of SVV, the invention provides methods of making new oncolytic virus derivatives and of altering viruses to have tumor-specific tropisms.

Description

[0001] This disclosure contains copyrighted material. [0002] The copyright owner has no objection to the reproduction by anyone of the patent document or patent disclosure appearing in the USPTO patent files or records, but objects to the retention of any and all copyright rights. [0003] All patent applications, published patent applications, and issued patents, texts, and references cited in this specification are hereby incorporated by reference in their entirety to more fully describe the state of the art to which this invention pertains. [0004] This application claims priority to U.S. Serial No. 60 / 506,182, filed September 26, 2003, which is hereby incorporated by reference in its entirety. Background of the invention [0005] Viral therapy holds great promise for cancer treatment. Oncolytic viruses, whether natural or engineered, that specifically bind and kill cancer cells are more effective and less toxic than selective treatments such as chemotherapy and radiati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C07H21/02C12N15/85C12N15/86C07K16/00C12Q1/70C12Q1/68A61KA61K35/76A61K35/768C07K14/085C12N7/04
CPCC12N7/00C07K16/00A61K35/768C12N2770/32032C07K14/005C12N2770/32022C12N2770/32061A61K35/76A61P35/00A61P43/00C12N15/11C07K14/085C12N15/86
Inventor 保罗·L·哈伦贝克卡尔·M·哈伊尚西·加内什塞希达·雷迪·波利斯徐玲杨敬平程诚
Owner NOVARTIS PHARM AG
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