Application of avobenzone in the preparation of antitumor drugs

An anti-tumor drug, avobenzone technology, applied in the field of medicine, can solve the problems of not verifying the therapeutic effect of avobenzone on skin cancer, researching and verifying the significance of skin cancer treatment, and not having a cell biology perspective, etc. Clinical drug resistance, improving application value, and clear effect of countermeasures for side effects

Active Publication Date: 2019-09-17
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 the preparation of antitumor drugs
  • Application of avobenzone in the preparation of antitumor drugs
  • Application of avobenzone in the preparation of 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 and 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 of NCBI database (NCBI accession number: NM_001281496.1), primers encoding PHB1 gene were designed and synthesized by Primer Premier 5.0, and double restriction sites were added at both ends Point (upstream end restriction site is BamH1, downstream end restriction site is EcoR1, both purchased from TAKARA), PCR amplification of the target gene band.

[0030] 2. Use 1% agarose gel electrophoresis to detect the PCR product, and use the DNA recovery kit (Tiangen Biochemical Technology Co., Ltd., Item No. DP214) to recover and purify the target fragment. Use endonuclease to double digest the target gene fragment and the empty vector pET (Biyuntia...

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, the co-immunoprecipitation experiment (Co-IP) was carried out in this example. The role of Zong is further studied. The specific operation steps of Co-IP are as follows:

[0072] (1) Two groups of 1 million colon cancer HCT116 cells in each group were plated in a cell culture dish in advance, and DMSO and avobenzone were added respectively. DMSO is added in the ratio of 1μL:1000μL DMEM culture solution, and Avobenzone is diluted with DMEM culture solution, and the final concentration is 10μM;

[0073] (2) Incubate 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 make the cells fall off the culture dish wall; then terminate with 5 mL of complete culture ...

Embodiment 3

[0082] Example 3 Study of MEK activation level

[0083] Protein MEK is a downstream molecule of PHB-c-Raf1 signaling pathway. By detecting the activation level of MEK, the phosphorylation state (pMEK), it can be confirmed whether the interaction between PHB and c-Raf1 is disrupted. The decrease of pMEK level also means that the metastatic ability of colon cancer cells is decreased.

[0084] Detect the activation level of MEK by western blot, the specific steps are as follows:

[0085] 1. Pave colon cancer cell HCT116 or colon cancer cell RKO (purchased from ATCC, article number CRL-2577) in a six-well plate with 500,000 cells per well. Add 2μL each of DMSO (control group) and avobenzone (experimental group) to the culture wells, and add 2000μL DMEM complete culture solution to each well. The final concentration of avobenzone is 10μM. Mix the culture medium and incubate the treated cells at 37°C for 48 hours.

[0086] 2. Wash the cells twice with PBS, add 0.5mL 0.25% trypsin to each ...

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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 particularly relates to the application of avobenzone in the preparation of antitumor drugs. Background technique [0002] Colon cancer is a common malignant tumor with complicated causes. Currently, there is no specific targeted drug to treat this type of cancer. Surgical resection, radiotherapy and chemotherapy are still the three major treatments for colon cancer. However, traditional treatment strategies have disadvantages such as long cycles, large side effects, and easy recurrence. Targeted drugs originally developed for the treatment of lung cancer, such as gefitinib, can also be used in the treatment of colon cancer. However, this kind of anti-cancer drugs that target a single gene or protein often develop resistance after a period of clinical use phenomenon. Therefore, it is of urgent practical significance to obtain safe, efficient, and long-lasting colon cancer targeted drugs. [0003...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/12A61P35/00A61P35/04
Inventor 何庆瑜杨杰李斌
Owner JINAN UNIVERSITY
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