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Striatin interacting protein inhibitor and application of inhibitor to preparation of anti-tumor drug

A protein inhibitor, anti-tumor drug technology, applied in the field of biomedicine

Pending Publication Date: 2019-03-12
HANGZHOU GONGCHU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It can be seen that STRIP plays an important role in the cell differentiation of lower and higher organisms, and is also related to the metastasis of breast cancer cells. However, whether there is a regulatory relationship between STRIP and the growth and inhibition of other tumors has not been reported yet.
In addition, when oncolytic vaccinia virus is used as a carrier to cooperate with STRIP, there is no related report on whether there is only a simple expression relationship between the two vectors, or whether STRIP can also regulate the replication of oncolytic vaccinia virus in tumor cells

Method used

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  • Striatin interacting protein inhibitor and application of inhibitor to preparation of anti-tumor drug
  • Striatin interacting protein inhibitor and application of inhibitor to preparation of anti-tumor drug
  • Striatin interacting protein inhibitor and application of inhibitor to preparation of anti-tumor drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1, siRNA interference of STRIP1 and STRIP2 leads to a significant decrease in the level of antiviral factors in malignant glioblastoma U87MG cells

[0047] siRNA of STRIP1 or STRIP2 (purchased from Santa Cruz Biotechnology) was transfected into glioma cells U87MG, and non-specific RNA (from Santa Cruz Biotechnology) was used as a control. The cell transfection procedure is as follows:

[0048] Operate according to the instructions of Attractene Transfection Reagent Handbook of QIAGEN Company:

[0049] (1) Before transfection, inoculate 3×10 per well in a 24-well plate 4 500 μL of DMEM medium containing 1% double antibody and 10% fetal bovine serum was added to each well. 37°C, 5% CO 2 Cultured in an incubator.

[0050] (2) After the cells adhere to the wall (generally 2-3 hours, depending on the situation of different cells), transfect, add an appropriate amount of RNA into a 0.5mL centrifuge tube according to the siRNA instructions.

[0051] (3) Add 1.5 μL...

Embodiment 2

[0056] Example 2, shRNA interference of STRIP1 down-regulates the expression level of antiviral factor IFI16 in liver cancer cells SMMC-7721

[0057] The shRNA interference plasmid of STRIP1 was transfected into SMMC-7721 liver cancer cells to establish a stable expression cell line. The non-specific RNA plasmid was used as a control to establish a control cell line. The plasmid transfection method was the same as that in Example 1. The transfected cells were screened with puromycin to establish a stably transfected cell line. The cell selection method was as follows:

[0058] (1) Add puromycin with different concentration gradients to the cells to be screened before screening.

[0059] (2) Generally, after 48 hours, observe which concentration gradient of puromycin has a cell survival rate of zero, and then use this concentration of puromycin to select stable transfected cells.

[0060] (3) Transfect the cells to be screened, and observe the transfection rate of the cells ...

Embodiment 3

[0089] Example 3. shRNA interference of STRIP1 can inhibit the activation of the interferon-stimulated response element ISRE in tumor cells caused by vaccinia virus infection

[0090] A plasmid for STRIP1 shRNA interference was constructed and transfected into U87MG cells to establish a stable expression cell line. A plasmid expressing non-specific RNA was used as a control, and a control cell line was established after transfecting cells. Two kinds of cells were infected or not infected with vaccinia virus, and transfected into the ISRE-Luc reporter plasmid, and the cell protein was collected for reporter gene detection, as follows:

[0091] Spread U87MG cells in good growth state on a 24-well plate with 80,000 cells per well and 0.5 mL of fresh culture medium. After the cells adhered to the wall, co-transfect the cells with ISRE firefly luciferase reporter gene plasmid and Renilla luciferase plasmid at a concentration of 500:1 (operate according to the above cell transfecti...

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Abstract

The invention relates to the technical field of biomedicine engineering, and provides an application of an STRIP (striatin interacting protein) inhibitor to preparation of an anti-tumor drug, a striatin interacting protein inhibitor recombinant vector, an application of the recombinant vector to preparation of the anti-tumor drug and an anti-tumor drug composition taking the striatin interacting protein inhibitor or the striatin interacting protein inhibitor recombinant vector as active components. The inhibitor for inhibiting a STRIP can be used for preparing the anti-tumor virus drug, particularly, the inhibitor and a vaccinia virus are recombined into the recombinant vector, the advantages of the STRIP inhibitor and the vaccinia virus can be combined, a new target spot is provided for viral therapy of tumors, and the STRIP inhibitor has a wide clinical application prospect.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of striatin interacting protein (STRIP) as a target in the preparation of antitumor drugs, a carrier and a drug containing it. Background technique [0002] Oncolytic viruses refer to therapeutically valuable viruses that can selectively infect and damage tumor tissues. Oncolytic viruses have a long history. So far, oncolytic viruses belonging to 10 virus families have entered clinical trials, including Adenovirus, Coxackie virus, Herpes simplex virus (HSV) ), Measles virus, Newcastle disease virus, Parvovirus, Poliovirus, Reovirus, Vaccinia virus and vesicular oral Vesicular stomatitis virus. Herpes virus T-VEC has been approved by the FDA for the treatment of melanoma. Overall, oncolytic viruses have shown considerable safety and efficacy in clinical trials. [0003] Vaccinia virus, a large double-stranded DNA virus in the poxvirus family, has several ad...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K35/768A61P35/00A61P35/04C12N15/63C12N15/113
CPCA61K48/005A61P35/00A61P35/04A61K35/768C12N15/113C12N15/63C12N2310/14C12N2710/24143C12N15/86Y02A50/30C12N2310/531C12N15/1137C12N2710/24132
Inventor 李恭楚
Owner HANGZHOU GONGCHU BIOTECH CO LTD
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