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Application of RORa protein and agonist thereof in preparation of acute DNA damage repairing agent

A DNA damage and agonist technology, applied in the field of biomedicine, can solve problems such as adverse reactions

Active Publication Date: 2022-08-05
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention intends to provide the application of RORa protein in the preparation of acute DNA damage repair agent, to solve the technical problem that a large number of adverse reactions will occur during tumor radiotherapy or chemotherapy

Method used

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  • Application of RORa protein and agonist thereof in preparation of acute DNA damage repairing agent
  • Application of RORa protein and agonist thereof in preparation of acute DNA damage repairing agent
  • Application of RORa protein and agonist thereof in preparation of acute DNA damage repairing agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The cholesterol sulfate compound (Cholesteryl sulfate, CS for short) used in this scheme is specifically cholesterol sulfate sodium salt (CAS: 2864-50-8), and the molecular formula is shown in formula (I).

[0056]

[0057] (1) Preparation of CS solution: CS was dissolved in DMSO to prepare a CS stock solution with a concentration of 50 mg / ml. CS working solution is formed by diluting CS stock solution to 5 mg / ml. CS working solution was used to prepare the test agent (injection) of CS group, and its components were: CS working solution 100 μL, Tween 80 20 μL and PBS 880 μL (PBS is conventional phosphate buffered saline solution in the prior art). At the same time, the control reagent (placebo) of NC group was prepared, and its components were: 100 μL of DMSO, 20 μL of Tween 80 and 880 μL of PBS.

[0058] (2) Experimental scheme:

[0059] This program uses C57BL6 mice (8-12 weeks old) for in vivo experiments. This mouse species is the conventional experimental mous...

experiment example 1

[0065] (1) Immunofluorescence detection (γH2AX)

[0066] Following γ-ray irradiation, certain DNA damage, including DNA double-strand breaks (DSBs), occurs in mouse cells. DNA damage activates the DNA damage response (DDR). Among them, DSB formation triggers the activation of many factors, including phosphorylation of the histone variant H2AX, resulting in γH2AX. Phosphorylation of H2AX plays a key role in the DDR and is required for the assembly of DNA repair proteins at sites containing damaged chromatin and the activation of checkpoint proteins that arrest cell cycle progression. Analysis of H2AX expression can be used to detect the genotoxic effects of different toxicants, and histone H2AX phosphorylation is the most prominent marker of DNA damage. In this protocol, γH2AX is used as a relevant indicator of DNA damage by immunofluorescence technology. The immunofluorescence experimental steps are roughly as follows: 5 Cell numbers were added dropwise on glass slides and ...

experiment example 2

[0071] Experimental example 2: RORa protein knockout experiment

[0072] The conventional Cre / loxP recombinase system in the prior art is used to knock out the mouse RORa protein, and then the aging phenotype of the mouse after the RORa protein deletion is detected. Cre / loxP-based gene knockout involves first introducing loxP sequences into the genome of embryonic stem cells, and then genetically modifying or changing the target gene through Cre-mediated recombination to obtain target gene knockout mice. The experimental results showed that knocking out RORa in the blood stem cells of young mice resulted in a decrease in the DNA repair capacity of the blood stem cells of young mice. Figure 8 Fluorescence immunoassay results of γH2AX in RORa knockout mice, Figure 9 is the statistical graph of γH2AX foci, Figure 10 Graph of the fluorescence intensity of γH2AX. Figure 8-Figure 10 The experimental results showed that DNA damage in mouse HSCs was aggravated after RORa gene k...

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Abstract

The invention relates to the field of biological medicines, relates to a DNA (Deoxyribose Nucleic Acid) damage repairing medicine, and in particular relates to application of RORa protein and an agonist thereof in preparation of an acute DNA damage repairing agent. According to the scheme, a large number of in-vivo experiments prove that under the condition that RORa protein is activated, cell acute DNA damage repair can be accelerated, especially DNA damage caused by gamma-ray irradiation can be effectively relieved, DNA self-repair of damaged cells can be accelerated, and the RORa protein can be applied to DNA repair protection after radiotherapy and has a protective effect on radioactive ray damage. The RORa protein agonist can solve the technical problem that a large number of adverse reactions occur in the tumor radiotherapy process, and then the radiotherapy effect is improved. The DNA repair scheme depending on the RORa protein is clear in action target, the RORa protein agonist is small in toxic effect, and the drug is safe and efficient and has good application prospects.

Description

technical field [0001] The invention relates to the field of biomedicine, relates to a DNA damage repair medicine, and particularly relates to the application of RORa protein and its agonist in the preparation of an acute DNA damage repair agent. Background technique [0002] Radiation therapy for tumors, or radiotherapy for short, is the use of radiation to treat cancer. Radiation therapy has experienced more than a century of development history. After Roentgen discovered X-ray and Marie Curie discovered radium, they were soon used in the clinical treatment of malignant tumors. Until now, radiotherapy is still an important local treatment method for malignant tumors. At present, about 70% of cancer patients need radiation therapy in the process of cancer treatment, and about 40% of cancers can be cured with radiation therapy. The role and status of radiotherapy in tumor treatment is increasingly prominent. Radiation includes alpha, beta, and gamma rays produced by radio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/575A61K9/00A61K47/26A61K47/02A61P35/00
CPCA61K31/575A61K9/0019A61K47/26A61K47/02A61P35/00Y02A50/30
Inventor 李宁王楠邱焕迪何巍邱强高珩寒尹月霞王勃川
Owner SICHUAN UNIV
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