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Compositions and method for determining anti-viral drug susceptibility and resistance and anti-viral drug screening

An antiviral drug and sensitivity technology, applied in the field of antiviral drug susceptibility and resistance testing, can solve problems such as the spread of retroviruses, achieve the effects of optimizing treatment plans, overcoming drug resistance problems, and effective treatment efficacy

Inactive Publication Date: 2006-07-12
病毒科学公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A major problem with the use of retroviruses is the possibility of spreading retroviruses capable of replicating

Method used

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  • Compositions and method for determining anti-viral drug susceptibility and resistance and anti-viral drug screening
  • Compositions and method for determining anti-viral drug susceptibility and resistance and anti-viral drug screening
  • Compositions and method for determining anti-viral drug susceptibility and resistance and anti-viral drug screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0185] Using segments containing patient origin and having replacement promoters

[0186] Non-functional indicator gene for resistance test vector HIV

[0187] Drug Sensitivity and Resistance Testing

[0188] Indicator gene viral vector - construction method

[0189] Design an indicator gene viral vector containing a non-functional indicator gene with a replacement promoter using HIV genomic and subgenomic viral vectors containing viral genes targeted by antiviral drugs. The indicator gene viral vectors pLG-lucPP and pCG-lucPP are based on the genomic viral vectors pLG and pCG; each carrying a deletion in the HIVenv gene. Resistance test vectors derived from the genomic indicator gene viral vectors pLG-lucPP and pCG-lucPP contain patient sequence acceptance sites for insertion of patient-derived segments and are packaged with expression of the amphipathic MLV 4070A env gene product carrier used together. The indicator gene viral vectors pLS...

Embodiment 2

[0235] Utilize segments containing patient origin and non-functional

[0236] HIV drug susceptibility and resistance testing with resistance test vectors for indicator genes

[0237] Indicator Gene Viral Vectors—Construction Method

[0238] Genomic indicator gene viral vectors pLG-lucPP-HS, pLG-lucPC-PB, pCG-lucPC-HS and pCG-lucPC-PB with the recipient site of the patient sequence and the resistance test vectors derived from them each at 5' to the 3' direction contains the following elements ( Figure 4B ): 1) HIV-LTR U3 region (pLG-lucPP-HS and pLG-lucPC-PB) or the first CMV IE enhancer-promoter (pCG-lucPC-HS and pCG-lucPC-PB), 2) HIV- LTR R and U5 regions, 3) coding regions of HIV gag-pol, vif, tat, rev, vpu, deleted env, and nef bases, 4) 5' coding containing luciferase gene inserted in deleted env gene region, 5) a second indicator cassette containing the 3' coding region of the luciferase gene inserted in the deleted 3' HIV-LTR U3 region, and 6) 3' HIV-LTR R and ...

Embodiment 3

[0250] Utilizes segments containing patient origin and has inverted introns

[0251] Non-functional indicator gene for resistance test vector HIV

[0252] Drug Sensitivity and Resistance Testing

[0253] Resistance test vectors containing non-functional indicator genes with inverted introns were designed using the HIV genome and subgenomic viral vectors containing the antiviral target genes described in Example 1.

[0254] Indicator Gene Viral Vector - Inverted Intron

[0255] Genomic indicator gene viral vectors pLG-lucII-HS, pLG-lucII-PB, pCG-lucII-HS and pCG-lucII-PB with patient sequence acceptance sites, and resistance test vectors derived from them, each at 5' to the 3' direction contains the following elements ( Figure 5B): 1) HIV-LTR U3 region (pLG-lucII-HS and pLG-lucII-PB) or the first CMV IE enhancer-promoter (pCG-lucII-HS and pCG-lucII-PB), 2) HIV- LTR R and U5 regions, 3) coding regions of HIV gag-pol, vif, vpr, tat, rev, vpu, ...

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Abstract

This invention provides a method for determining susceptibility for an anti-viral drug comprising: (a) introducing a resistance test vector comprising a patient-derived segment and an indicator gene into a host cell; (b) culturing the host cell from (a); (c) measuring expression of the indicator gene in a target host cell; and (d) comparing the expression of the indicator gene from (c) with the expression of the indicator gene measured when steps (a)-(c) are carried out in the absence of the anti-viral drug, wherein a test concentration of the anti-viral drug is present at steps (a)-(c); at steps (b)-(c); or at step (c). This invention also provides a method determining anti-viral drug resistance in a patient. This invention also provides a method for evaluating the biological effectiveness of a candidate anti-viral drug compound. Compositions including resistance test vectors comprising a patient-derived segment and an indicator gene and host cells transformed with the resistance test vectors are provided.

Description

technical field [0001] The present invention relates to antiviral drug susceptibility and resistance tests for identifying effective drug modalities for treating viral infections. The present invention further relates to novel vectors, host cells and compositions useful for conducting susceptibility and resistance tests to these novel antiviral drugs. The present invention also relates to screening drug candidates for their ability to inhibit selected viral sequences and / or viral proteins. More specifically, the present invention relates to the use of recombinant DNA technology to first construct a resistance test vector containing patient-derived segments and indicator genes, then introduce the resistance test vector into host cells, and determine the target host cells in the presence of antiviral drugs Expression and repression of indicated gene products in . The present invention also relates to means and methods for identifying antiviral drugs that have a distinct resist...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12N15/49C12N15/64C12N15/09C12N5/00C12N5/10C12P21/02C12Q1/66C12Q1/68C12R1/91G01N33/50
CPCG01N2800/52G01N33/5008C12Q1/6897G01N33/5067G01N33/5044G01N33/505G01N33/502C12Q1/02C12N15/11C12N15/64
Inventor 丹尼尔·J·卡彭克里斯托·约翰·彼得罗普洛斯
Owner 病毒科学公司