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Clonal strains of attenuated vaccinia viruses and methods of use thereof

A virus strain, virus strain technology, applied in the direction of viruses, antiviral agents, viruses/phages, etc., can solve problems such as reducing and weakening the oncolytic properties of viruses

Inactive Publication Date: 2014-10-08
GENELUX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, methods of attenuating the virus can reduce or attenuate the oncolytic properties of the virus

Method used

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  • Clonal strains of attenuated vaccinia viruses and methods of use thereof
  • Clonal strains of attenuated vaccinia viruses and methods of use thereof
  • Clonal strains of attenuated vaccinia viruses and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0071] 4. Preparation of the virus

[0072] F. Pharmaceutical compositions, combinations and kits

[0073] 1. Pharmaceutical composition

[0074] 2. Host cell

[0075] 3. Combination

[0076] 4. Kit

[0077] G. Methods of treatment, diagnosis and monitoring

[0078] 1. Treatment

[0079] 2. Diagnosis and monitoring methods

[0080] 3. Application

[0081] a. Steps before applying the virus

[0082] b. Application mode

[0083] c. Dosage and dosing schedule

[0084] d. Co-administration

[0085] i. Application of multiple viruses

[0086] ii. Therapeutic compounds

[0087] iii. Immunotherapy and biological therapy

[0088] e. Object status

[0089] 4. Monitoring

[0090] a. Monitoring viral gene expression

[0091] b. Monitor tumor size

[0092] c. Monitor antibody titer

[0093] d. Monitoring general health diagnosis

[0094] e. Monitoring co-treatment

[0095] H. Examples

[0096] A. Definition

[0097] Unless otherwise specified, all technical and academic terms used in the present invention have the same...

Embodiment 1

[0564] Example 1: Isolation of LIVP clone isolate

[0565] Plate African green monkey kidney fibroblasts CV-1 cells (ATCC No. CCL-70; American Type Culture Collection (Manassas, VA)) on a 6-well plate with a density of 5×10 5 Cells / well, in Dulbecco's modified Eagle's medium (DMEM) supplemented with 1% antibiotic-antifungal solution (Mediatech, Inc., Herndon, VA) and 10% fetal bovine serum (FBS, Mediatech, Inc., Herndon, VA) , Mediatech, Inc., Herndon, VA) 5% CO at 37℃ 2 Grow overnight in a humidified incubator. When the cells reach 90% confluence, CV-1 cells are diluted 10-fold with vaccinia virus LIVP (vaccinia virus strain, originally produced by adapting Lister strain (ATCC Catalog No.VR-1549) on calf skin) (Moscow Institute of Virus Preparation, Russia, Al'tshtein et al., (1983) Dokl. Akad. Nauk USSR285: 696-699)) infection. Use serial dilutions to infect to ensure the separation of fully separated plaques. Two days after infection, 8 well-separated plaques that showed a l...

Embodiment 2

[0566] Example 2: In vitro infection of various normal and cancer cell lines

[0567] Analyze the growth of LIVP clone isolates in various normal and cancer cell types, and compare them with the parent LIVP strain, two LIVP-derived strains GLV-1h68 (see U.S. Patent No. 7,588,767) and GLV-1h74 (see U.S. Patent Application Publication No. 2009-0098529 and 2009-0053244) and the growth of vaccinia virus WR (ATCC Catalog No. VR-1354) were compared. GLV-1h68 contains a DNA insertion in the locus of the LIVP strain (SEQ ID NO: 9). GLV-1h68 contains expression cassettes encoding detectable marker proteins at F14.5L (also called F3 in LIVP), thymidine kinase (rK) and hemagglutinin (HA) loci. Contains early / late promoter P which is synthesized by vaccinia virus SEL ((P SEL ) Ruc-GFP)-controlled Ruc-GFP cDNA molecule (a fusion of DNA encoding Renilla luciferase and DNA encoding GFP) is inserted into the F14.5L locus; contains the early / late start of vaccinia virus Child P 7.5k ((P 7.5k ...

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Abstract

Clonal strains of vaccinia viruses are provided. Also provided are methods of identifying and isolating attenuated and oncolytic clonal strains from virus preparations, Modified recombinant forms of the clonal strains also are provided. The clonal strains and virus preparations can be used for diagnostic and therapeutic methods, in particular for therapy and diagnosis or monitoring treatment of proliferative disorders, including neoplastic diseases, such as, but are not limited to, solid tumors and blood cancers.

Description

[0001] Related application [0002] This application claims priority to the U.S. Provisional Application Serial No. 61 / 517,297 filed on April 15, 2011 and the U.S. Provisional Application Serial No. 61 / 628,684 filed on November 4, 2011. Each application belongs to Aladar A. Szalay, Nanhai Chen, Yong A. Yu and Qian Zhang and the invention is titled "Clonal Strains of Attenuated Vaccinia Viruses and Methods of Use Thereof". [0003] This application is related to the U.S. Patent Application No. (Attorney Dkt. 61 / 628,684 priority. [0004] When permitted, the subject matter of each of the aforementioned applications is incorporated herein in its entirety. [0005] Introduce the sequence listing in electronic form [0006] The electronic version of the sequence table is submitted here, and its contents are all incorporated into this article. The size of the electronic file is 4.44Mb and the name is 4832seqPC1.txt. Invention field [0007] Vaccinia virus isolate. Background of the inventi...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/768
CPCA61K35/768C12N2710/24121C07K16/22C12N2710/24143C12N2710/24132C07K2317/622C12N7/00A61P1/04A61P1/18A61P11/00A61P13/08A61P15/00A61P31/12A61P35/00A61P35/04A61P43/00
Inventor A·A·绍洛伊N·G·陈Y·A·于Q·张
Owner GENELUX
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