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Application of IAP inhibitor and oncolytic virus in preparation of antitumor drug

An oncolytic virus and anti-tumor technology, applied in the field of biomedicine

Active Publication Date: 2017-01-04
GUANGZHOU VIROTECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the patent 201510990705.7 previously applied by the inventor, chrysophanol and its derivatives are used as anti-tumor synergists of M1 virus. 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 IAP inhibitor and oncolytic virus in preparation of antitumor drug
  • Application of IAP inhibitor and oncolytic virus in preparation of antitumor drug
  • Application of IAP inhibitor and oncolytic virus in preparation of antitumor drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 LCL161 and M1 virus significantly increase the morphological lesions of human colorectal cell carcinoma lines

[0059] Material:

[0060] Human colorectal carcinoma HCT116, M1 virus, high glucose DMEM medium, inverted phase contrast microscope.

[0061] method:

[0062] a) Cell culture: human colorectal cancer HCT116 was grown in DMEM complete medium containing 10% FBS, 100U / ml penicillin and 0.1mg / 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.

[0063] 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 a cell sus...

Embodiment 2

[0066] Example 2 The combined treatment of LCL161 / Birinapant and M1 virus significantly reduced the survival rates of human colorectal cell carcinoma lines and human hepatocellular carcinoma lines;

[0067] Material:

[0068] Human colorectal cancer HCT116, SW480, human hepatocellular carcinoma Huh7, PLC, M1 virus, high glucose DMEM medium, automatic enzyme-linked detection microplate reader.

[0069] method:

[0070] 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 adhered to the wall. The experiment was divided into control group, single drug group (5μM LCL161 or Birinapant alone), M1 infection group and LCL161 / M1 or Birinapant / M1 combination group. The doses used were: M1 virus infected cells, and set different MOI...

Embodiment 3

[0075] Example 3 Inhibition of cIAP1 and cIAP2 in synergy with the anti-tumor effect of M1 oncolytic virus

[0076] Material:

[0077] M1 virus, human colorectal cancer cell HCT116, liver cancer cell Huh7, RNA interference fragments of cIAP1 and cIAP2, MTT (methyl azolazolium blue), phase contrast microscope.

[0078] cIAP1 Interfering Fragment (Si RNA):

[0079] Interference fragment 1:

[0080] Sense strand (SEQ ID NO.1)

[0081] 5'-GUGAGUUCUUGAUACGAAUdTdT-3'

[0082] Antisense strand (SEQ ID NO.2)

[0083] 5'-AUUCGUAUUCAAGAACUCACdTdT-3'

[0084] Interference fragment 2:

[0085] Sense strand (SEQ ID NO.3)

[0086] 5'-GAAUACGUCUCCAAUGAGAdTdT-3'

[0087] Antisense strand (SEQ ID NO.4)

[0088] 5'-UCUCAUUGGAGACGUAUUCdTdT-3'

[0089] Interference fragment 3:

[0090] Sense strand (SEQ ID NO.5)

[0091] 5'-GCAAGUGCUGGAUUCUAUUUdTdT-3'

[0092] Antisense strand (SEQ ID NO.6)

[0093] 5'-AAUAGAAUCCAGCACUUGCdTdT-3'

[0094] cIAP2 Interfering Fragment (Si RNA):

[009...

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Abstract

The invention belongs to the field of biological medicines and relates to an application of combination of a Caspase activator and an oncolytic virus in preparation of an antitumor drug. The condition that the antitumor effect of the oncolytic virus can be improved with the Caspase activator is discovered for the first time, the combination of the Caspase activator and the oncolytic virus has a quite high synergistic effect, and an effective therapeutic schedule is provided for oncotherapy with low drug sensitivity.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of the combination of Caspase activator 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 harmless to normal cells. [0003] M1 virus (Alphavirus M1) belongs to the genus Alphavirus (Alphavirus), and it has a relatively good application effect in the preparation of antitumor drugs. For example, the Chinese invention patent application 201410425510.3 discloses that the M1 virus can selectively cause tumor cell death ...

Claims

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Application Information

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IPC IPC(8): A61K35/768A61K45/06A61K31/427A61K31/404A61P35/00
CPCA61K31/404A61K31/427A61K35/768A61K45/06A61K2300/00A61P35/00A61K31/713A61K9/0019A61K9/19A61K9/48A61K9/7023
Inventor 蔡静邓静张海鹏吕凌林园W.K.凯夫尼高光坪程世源林穗珍龚守芳胡骏白雪涛肖晓李凯梁剑开谭亚倩朱文博银巍颜光美
Owner GUANGZHOU VIROTECH PHARMA
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