Application of adefovir dipivoxil in preparing antitumor drugs

A technology of adefovir dipivoxil and anti-tumor drugs, which is applied in the field of medicine, can solve the problem of the influence and effect of adefovir dipivoxil on the growth of cancer cells, and achieve the purpose of inhibiting the growth, division and proliferation of colon cancer cells , moderately priced effect

Active Publication Date: 2018-12-07
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 preparing antitumor drugs
  • Application of adefovir dipivoxil in preparing antitumor drugs
  • Application of adefovir dipivoxil in preparing antitumor drugs

Examples

Experimental program
Comparison scheme
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 an application of adefovir dipivoxil in preparing antitumor drugs. The invention first finds a brand-new action mechanism of adefovir dipivoxil, namely, adefovir dipivoxil is capable of obviously damaging interaction between proteins KCTD12 and CDK1, so as to damage a tumor cell signaling pathway and restrain the growth of tumor cells. Adefovir dipivoxil has high specificityto targeting tumor, and meanwhile, adefovir dipivoxil acts on the interaction of two proteins, so that occurrence probability of drug resistance is extremely reduced. The invention further first proves that adefovir dipivoxil is capable of obviously restraining fission and multiplication of colon cancer cells through a molecular biological test, a cytology test and an animal test, the applicationscope of adefovir dipivoxil is widened and the application value of adefovir dipivoxil is increased. Meanwhile, adefovir dipivoxil is a drug widely applied to market and clinic, has few side effects,has clear solutions for the side effects, is in moderate cost and has a 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 Applications(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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