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Application of adefovir dipivoxil in the preparation of antitumor drugs

An anti-tumor drug, the technology of adefovir dipivoxil, applied in the field of medicine, can solve problems such as the influence and effect of adefovir dipivoxil on the growth of cancer cells that have not yet been seen, and achieves inhibition of the growth of colon cancer cells, high specificity, and high specificity. Avoid the effects of drug resistance

Active Publication Date: 2021-02-05
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no relevant reports on the influence and effect of adefovir dipivoxil on the growth of cancer cells (especially colon cancer cells)

Method used

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  • Application of adefovir dipivoxil in the preparation of antitumor drugs
  • Application of adefovir dipivoxil in the preparation of antitumor drugs
  • Application of adefovir dipivoxil in the preparation of antitumor drugs

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Study the anti-tumor mechanism of adefovir dipivoxil by superELISA technology

[0030] 1. Preparation of fusion protein KCTD12-his, CDK1-gst

[0031] (1) Expression and purification of KCTD12-his fusion protein

[0032] 1. Construction of the recombinant expression vector pET-KCTD12: According to the human KCTD12 protein gene sequence in the NCBI database (NCBI accession number: NM_138444.3), use Primer Premier 5.0 to design and synthesize primers encoding the KCTD12 gene, and add double restriction sites at both ends (The restriction site at the upstream end is BamH1, and the restriction site at the downstream end is EcoR1, both purchased from TAKARA Company), and the target gene band is amplified by PCR.

[0033] 2. The PCR product was detected by 1% agarose gel electrophoresis, and the target fragment was recovered and purified using a DNA recovery kit (Tiangen Biochemical Technology Co., Ltd., catalog number: DP214). Use endonucleases to double-digest th...

Embodiment 2

[0072] Example 2 Co-immunoprecipitation experiment

[0073] Co-immunoprecipitation experiment (Co-IP) is an important means to test the interaction of proteins in vivo. In order to further study and verify the anti-cancer mechanism and effect of adefovir dipivoxil, the present embodiment carries out co-immunoprecipitation experiment (Co-IP) on Adefovir dipivoxil. Fuvir dipivoxil for further study.

[0074] In advance, 4 groups of 1 million colon cancer HCT116 (purchased from ATCC; product number: CCL-247) or HT29 cells (purchased from ATCC, product number: HTB-38) were spread in cell culture dishes in each group, and DMSO was added respectively, Drug #1, Drug #2, Drug #3. DMSO was added according to the ratio of 1 μL: 1000 μL DMEM medium, drugs 1 to 3 were diluted with DMEM medium, and the final concentration was 10 μM; the treated cells were cultured at 37°C for 24 hours, and then the cells were washed twice with PBS. Add 1 mL of 0.25% trypsin to digest the cells to make th...

Embodiment 3

[0083] Embodiment 3 Adefovir dipivoxil on the influence of cancer cell division

[0084] 1. Spread the colon cancer cells HCT116 in a 6-well plate, with 500,000 cells per well. In the experimental group, 2 μL of 10 mM adefovir dipivoxil was added to 2 mL of complete culture solution to make the final concentration 10 μM; in the control group, 2 μL of DMSO was added to 2 mL of complete culture solution. Incubate at 37°C for 48 hours.

[0085] 2. The cells were digested and counted, and 500,000 cells were taken from each of the experimental group and the control group. Wash the cells with PBS, wash away the attached culture medium, and centrifuge at 300g for 5 minutes.

[0086] 3. Prepare a mixture of 600 μL PBS and FBS in advance, the preparation method is 540 μL PBS+60 μL FBS, mix well and pre-cool on ice. Take 300 μL of the mixture of PBS and FBS to resuspend the cells, then add 700 μL of pre-cooled 100% ethanol to each group, and rotate at 4°C overnight (16-18 hours) for ...

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Abstract

The invention provides the application of adefovir dipivoxil in the preparation of antitumor drugs. The present invention discovers for the first time a brand-new mechanism of action of adefovir dipivoxil, which can significantly destroy the interaction between the protein KCTD12 and CDK1, thereby destroying the tumor cell signaling pathway, and then inhibiting the growth of tumor cells, with high specificity for targeting tumors; At the same time, since adefovir dipivoxil acts on the interaction between the two proteins, the emergence of drug resistance is avoided to the greatest extent. The present invention further confirms for the first time that adefovir dipivoxil can significantly inhibit the division and proliferation of colon cancer cells through molecular biology, cytology and zoology experiments, expands the application range of adefovir dipivoxil, and improves its application value. At the same time, adefovir dipivoxil is a drug widely used in the market and clinical practice, with less toxic and side effects, relatively clear countermeasures for side effects, and moderate price, which has important practical significance.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular to the application of adefovir dipivoxil in the preparation of antitumor drugs. Background technique [0002] Traditional chemotherapeutic drugs such as cisplatin drugs have relatively large toxic and side effects because they cannot specifically kill tumor cells. Most of the existing anti-tumor targeted drugs are drugs that target a single oncogene or a single protein. This type of drug can specifically recognize tumor cells and has a good therapeutic effect. Although this type of targeted drug can also be used to treat cancer, the targeted gene is prone to drug-resistant mutations under the continuous action pressure of this type of drug. Drug resistance, the mutated tumor cells lose their sensitivity to drugs and are no longer inhibited by drugs, resulting in tumor recurrence, that is, the existing targeted drugs can no longer work to inhibit tumor development. Therefore, it i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/675A61P35/00
CPCA61K31/675A61P35/00
Inventor 何庆瑜杨杰李斌
Owner JINAN UNIVERSITY
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