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Application of Bcl-xL inhibitor and oncolytic virus to preparation of anti-tumor medicines

An oncolytic virus and inhibitor technology, which is applied in the field of biomedicine to achieve the effects of increasing anti-tumor effect, enhancing inhibitory effect and improving therapeutic effectiveness

Active Publication Date: 2016-12-07
GUANGZHOU VIROTECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the patent 201510990705.7 previously applied by the inventor, chrysophanol and its derivatives are used as anti-tumor synergists of oncolytic viruses. The combination of the two can reduce the survival rate of tumor cells to 39.6%, but there is a big difference in its anti-cancer strength. room for improvement

Method used

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  • Application of Bcl-xL inhibitor and oncolytic virus to preparation of anti-tumor medicines
  • Application of Bcl-xL inhibitor and oncolytic virus to preparation of anti-tumor medicines
  • Application of Bcl-xL inhibitor and oncolytic virus to preparation of anti-tumor medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 ABT-263 and M1 virus significantly increase the morphological lesions of human hepatocellular carcinoma lines

[0050] Material:

[0051] Human hepatocellular carcinoma Hep3B, M1 virus, high glucose DMEM medium, inverted phase contrast microscope.

[0052] method:

[0053] a) Cell culture: Human hepatocellular carcinoma Hep3B was grown in DMEM complete medium containing 10% FBS, 100 U / ml penicillin and 0.1 mg / ml streptomycin; all cell lines were placed in 5% CO 2 , cultured and subcultured in a closed incubator with a constant temperature of 37°C (95% relative humidity), and observed the growth with an inverted microscope. The cells were subcultured every 2-3 days, and the cells in the logarithmic growth phase were taken for formal experiments.

[0054] b) Cell treatment and morphological observation: cells in the logarithmic growth phase were selected, and DMEM complete culture medium (containing 10% fetal bovine serum, 1% double antibody) was used to make...

Embodiment 2

[0057] Example 2 Combined treatment of ABT-263 or ABT-737 with M1 virus significantly reduces the survival rate of human cancer cell lines

[0058] Material:

[0059] Human hepatocellular carcinoma Hep3B, human bladder carcinoma T24, human colorectal carcinoma LoVo, M1 virus, high glucose DMEM medium, automatic enzyme-linked detection microplate reader.

[0060] method:

[0061] a) Cell inoculation, administration treatment: select logarithmic growth phase cells, DMEM complete culture medium (containing 10% fetal calf serum, 1% double antibody) to make cell suspension, with 4 × 10 3 The density per well was seeded in a 96-well culture plate. After 12 hours, the cells were completely attached to the wall. The experiment was divided into control group, ABT-263 alone group, M1 infection group and ABT-263 / M1 combination group or ABT-737 / M1 combination group. The dose used is: M1 virus infects cells; different dose gradients are set for M1 virus.

[0062] b) Reaction of MTT wit...

Embodiment 3

[0067] Example 3 Inhibition of Bcl-xL synergistic anti-tumor effect of M1 oncolytic virus

[0068] Material:

[0069] M1 virus, human liver cancer cell Hep3B, bladder cancer cell T24, Bcl-xL and Bcl-w RNA interference fragments, MTT (methyl azolium blue), phase contrast microscope.

[0070] Bcl-xL interference fragment (Si RNA):

[0071] Sense strand (SEQ ID NO.1)

[0072] 5'-GGAUACAGCUGGAGUCAGUdTdT-3'

[0073] Antisense strand (SEQ ID NO.2)

[0074] 5'-ACUGACUCCAGCUGUAUCCdTdT-3';

[0075] Bcl-w interference fragment (Si RNA):

[0076] Sense strand (SEQ ID NO.3)

[0077] 5'-CAGCUGUAUUCCAUUACAUdTdT-3'

[0078] Antisense strand (SEQ ID NO.4)

[0079] 5'-AUCUAAUGGAAUACAGCUGdTdT-3'

[0080] method:

[0081] Select the cells in the logarithmic growth phase, prepare the cell suspension with DMEM complete culture medium, and use 1×10 5 seeded in 6-well plates. After 24 hours, add liposome-encapsulated Si RNA target gene fragments and infect M1 virus 24 hours later. 48 ho...

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Abstract

The invention belongs to the field of biological medicines, and relates to application of Bcl-xL inhibitor and oncolytic virus to preparation of anti-tumor medicines. It is found for the first time that Bcl-xL inhibitor can be used for preparing oncolytic virus anti-tumor synergist. The invention meanwhile relates to a pharmaceutical composition containing Bcl-xL inhibitor and oncolytic virus, a medicine suit containing Bcl-xL inhibitor and oncolytic virus, and application of Bcl-xL inhibitor and oncolytic virus to treatment of tumors, especially tumors not sensitive to oncolytic virus.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of the combination of Bcl-xL inhibitor and oncolytic virus in the preparation of antitumor drugs. Background technique [0002] Oncolytic virus is a kind of replication-competent virus that targets and infects and kills tumor cells without destroying normal cells. Oncolytic virus therapy (oncolytic virotherapy) is an innovative tumor-targeted treatment strategy, which uses natural or genetically engineered viruses to selectively infect tumor cells and replicate in tumor cells to achieve targeted dissolution, The effect of killing tumor cells, but no damage to normal cells. [0003] M1 virus (Alphavirus M1) belongs to the genus Alphavirus (Alphavirus), and it has a good application effect in the preparation of antitumor drugs. For example, Chinese invention patent application 201410425510.3 discloses that M1 virus can selectively cause tumor cell death without affecting n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K35/768A61K45/06A61P35/00
CPCA61K35/768A61K45/00A61K45/06A61K2300/00A61K31/11A61K31/352A61K31/428A61K31/4725A61K31/5377A61K31/635A61P35/00A61K31/7105
Inventor 谭亚倩邓静林园张海鹏W.K.凯夫尼高光坪程世源吕凌林穗珍龚守芳胡骏白雪涛肖晓李凯梁剑开蔡静朱文博银巍颜光美
Owner GUANGZHOU VIROTECH PHARMA
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