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Application of avobenzone in preparing antitumor drugs

An anti-tumor drug, the technology of avobenzone, applied in the field of medicine, can solve the problems of researching and verifying the significance of skin cancer treatment, without the perspective of cell biology, and without verifying the therapeutic effect of avobenzone on skin cancer, so as to reduce the Clinical drug resistance, side effect response measures are clear, and the effect of improving application value

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

However, except for the function of sun protection, other effects of avobenzone, such as cancer treatment, have not been directly studied or reported. The reports on the prevention of skin cancer by avobenzone in the prior art are mainly discussed from the perspective of molecular biology. The principle of avobenzone in the prevention of skin cancer is that the drug decomposes under ultraviolet light to offset the energy of ultraviolet light, thereby reducing the damage of ultraviolet light to the skin. However, this literature does not verify the treatment of avobenzone for skin cancer that has occurred The effect of this drug has not been studied and verified from the perspective of cell biology (Reis JS, Corrêa MA, Chung MC, Dos Santos JL. Synthesis, antioxidant and photoprotection activities of hybrid derivatives useful to prevent skin cancer. Bioorg MedChem 2014;22(9):2733-8.)
[0005] At present, there is no prior art report on the influence and effect of avobenzone on the growth of cancer cells (especially colon cancer cells)

Method used

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  • Application of avobenzone in preparing antitumor drugs
  • Application of avobenzone in preparing antitumor drugs
  • Application of avobenzone in preparing antitumor drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Research on the Anti-tumor Mechanism of Avobenzone by SuperELISA Technology

[0027] 1. Preparation of fusion protein PHB1-his, C-Raf1-gst

[0028] (1) Expression and purification of PHB1-his fusion protein

[0029] 1. Construction of the recombinant expression vector pET-his: according to the human PHB1 protein gene sequence in the NCBI database (NCBI accession number: NM_001281496.1), use Primer Premier 5.0 to design and synthesize primers encoding the PHB1 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.

[0030] 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 the target...

Embodiment 2

[0070] Example 2 Co-immunoprecipitation experiment

[0071]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 avobenzone, this embodiment carried out co-immunoprecipitation experiment (Co-IP) on avobenzone The role of Zong should be further studied. The specific operation steps of Co-IP are as follows:

[0072] (1) Spread 2 groups of 1 million colon cancer HCT116 cells in a cell culture dish in advance, and add DMSO and avobenzone respectively. DMSO was added according to the ratio of 1 μL: 1000 μL DMEM culture solution, avobenzone was diluted with DMEM culture solution, and the final concentration was 10 μM;

[0073] (2) Cultivate the treated cells at 37°C for 24 hours, then wash the cells twice with PBS, add 1 mL of 0.25% trypsin to each culture dish to digest the cells to make them fall off the wall of the culture dish; then terminate with...

Embodiment 3

[0082] Example 3 MEK activation level research

[0083] The protein MEK is a downstream molecule of the PHB-c-Raf1 signaling pathway, and whether the interaction between PHB and c-Raf1 is destroyed can be confirmed by detecting the activation level of MEK, that is, the phosphorylation state (pMEK). Reduced levels of pMEK also imply reduced metastatic ability of colon cancer cells.

[0084] The activation level of MEK was detected by western blot method, and the specific steps were as follows:

[0085] 1. Spread colon cancer cells HCT116 or colon cancer cells RKO (purchased from ATCC, Cat. No. CRL-2577) in a six-well plate, with 500,000 cells per well. Add 2 μL each of DMSO (control group) and avobenzone (experimental group) to the culture wells, add 2000 μL of DMEM complete culture solution to each well, and the final concentration of avobenzone is 10 μM. Mix the culture solution and incubate the treated cells at 37°C for 48 hours.

[0086] 2. Wash the cells twice with PBS,...

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Abstract

The invention provides an application of avobenzone in preparing antitumor drugs. The invention first finds that avobenzone is capable of obviously damaging interaction between PHB and c-Raf1 two proteins, so as to damage a tumor cell signaling pathway and restrain the growth of tumor cells. Avobenzone has high specificity to targeting tumor, few side effects and clear solutions for the side effects. Avobenzone does not target to a single gene or protein, so that avobenzone will not generate lasting drug pressure resulting in mutation to a specific molecule or single protein in tumor cells, and occurrence probability of clinic drug resistance is greatly reduced. The invention further first proves that avobenzone is capable of obviously restraining colon cancer cell metastasis through a molecular biological test and a cytology test, the application scope of avobenzone is widened, the application value of avobenzone is increased and avobenzone has a practical significance in oncotherapy.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the application of avobenzone in the preparation of antitumor drugs. Background technique [0002] Colon cancer is a common malignant tumor with complex causes. Currently, there is no specific targeted drug to treat this type of cancer. Surgical resection, radiotherapy, and chemotherapy are still the three major means of colon cancer treatment. However, traditional treatment strategies have disadvantages such as long cycle, large side effects, and easy recurrence. Targeted drugs originally developed for the treatment of lung cancer, such as gefitinib, can also be used clinically for the treatment of colon cancer. However, anticancer drugs that target a single gene or protein often develop drug resistance after a period of clinical use Phenomenon. Therefore, it is of urgent practical significance to obtain safe, efficient and durable colon cancer targeted drugs. ...

Claims

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Application Information

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IPC IPC(8): A61K31/12A61P35/00A61P35/04
Inventor 何庆瑜杨杰李斌
Owner JINAN UNIVERSITY
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