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Preparation for combined treatment of head and neck neoplasms, and uses thereof

A head and neck tumor and preparation technology, applied in the field of biomedicine, can solve problems such as undiscovered head and neck tumors, and achieve the effects of reducing treatment cost, enhancing curative effect, and reducing dosage

Inactive Publication Date: 2015-08-05
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] According to the literature search, so far, no arginase has been found to induce autophagy of head and neck tumor cells, and a compound drug or composition composed of an autophagy inhibitor and arginase has not been found to be administered in combination or sequentially. Related reports on the treatment of head and neck tumors

Method used

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  • Preparation for combined treatment of head and neck neoplasms, and uses thereof
  • Preparation for combined treatment of head and neck neoplasms, and uses thereof
  • Preparation for combined treatment of head and neck neoplasms, and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Preparation of human recombinant arginase.

[0050] The coding sequence of human arginase (NM_000045.3) was obtained by PCR method. The full-length coding sequence of the gene is 969bp, with restriction sites XhoI and EcoRI. Use XhoI and EcoRI endonucleases to double-digest the PCR product and the empty plasmid vector respectively, connect the target gene to the vector and transform the ampicillin resistance plate, pick several positive transformants, take one of them to amplify and extract the plasmid, and carry out XhoI And EcoRI endonuclease double digestion identification. The recombinant plasmid was extracted, the recombinant vector was digested with BglⅡ to linearize, and Pichia pastoris was transformed by electroporation. Pick positive transformants. Through RDB, MM and MD plate screening, clones with the phenotype of his+muts were obtained, and the expression was further induced on the shaker level, and the expression of arginase on the shaker leve...

Embodiment 2

[0054] Embodiment 2: the prescription of autophagy inhibitor drug

[0055] Chloroquine (CQ), purchased from sigma company;

[0056] Ammonium chloride (NH 4 Cl), purchased from sigma company;

[0057] Hydroxychloroquine, purchased from sigma company;

[0058] 3-methyladenine (3-MA), purchased from sigma company;

[0059] PI3K inhibitor LY294002, purchased from Cell Signaling Technology;

[0060] Bafilomycin A1 (Bafilomycin A1), purchased from sigma company;

[0061] (1) Preparation of chloroquine: get appropriate amount of chloroquine dissolved in pure water to make 10mmol / L stock solution, filter and sterilize with a filter of 0.1 μm and store at 4°C, use PRMI-1640 medium for in vitro experiments (Thermo Fisher) Seoul Technology Company) was diluted 500-1000 times to inhibit autophagy.

[0062] (2) Preparation of ammonium chloride: Dissolve an appropriate amount of ammonium chloride in water to prepare a 0.4 mol / L storage solution, filter and sterilize with a 0.1 μm filt...

Embodiment 3

[0068] Example 3: Human recombinant arginase can induce autophagy in laryngeal carcinoma Tu212 cells

[0069] Laryngeal cancer Tu212 cells (purchased from the Cell Resource Center of the Central Laboratory of Hunan Xiangya Medical College) were treated with 0.12 U / ml arginase for 24 hours, then embedded in paraffin, sectioned, and stained, and the subunits of the cells were observed under a transmission electron microscope. Microstructure, the results are as figure 1 As shown in A, there are a large number of electron-dense inclusions in the cells of the administration group, and the typical double-membrane structure autophagosomes can be clearly seen after zooming in, while none were found in the control group.

[0070] Laryngeal carcinoma Tu212 cells were treated with 0.12U / ml arginase for 24h and 48h, and Cyto- Autophagy Detection Kit Autophagosome Staining Kit (ENZO Life Science Research Institute) was processed according to the instructions, and then the green fluoresce...

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Abstract

The present invention relates to the technical field of biomedicine. According to the present invention, research results show that arginase can significantly induce head and neck neoplasm cells to produce autophagy while the autophagy can partially counteract the anti-tumor effect of the arginase; and with use of the autophagy inhibitor to inhibit the autophagy, and the sensitivity of the head and neck neoplasm cells on the arginase can be increased so as to enhance the treatment effect of the arginase on the head and neck neoplasms. The present invention provides a preparation for combined treatment of head and neck neoplasms, specifically a drug composition comprising the autophagy inhibitor and the arginase. The present invention further provides applications of the drug composition in preparation of drugs for treatment of head and neck neoplasms, especially laryngeal cancer, hypopharyngeal cancer and nasopharyngeal cancer.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and is a preparation for combined treatment of head and neck tumors. Specifically, the present invention relates to a combined preparation of an autophagy inhibitor and arginase, and preparation of a drug for treating head and neck tumors. in the application. Background technique [0002] Arginase is a key enzyme involved in the urea cycle in the mammalian liver, where it converts arginine into ornithine and urea. Arginine is an essential amino acid to maintain the growth of some tumor cells. When the level of arginine in tumor cells is lower than a certain value, it will lead to cell death. Arginase can specifically and efficiently degrade arginine in vitro and in vivo. In vitro experiments, arginase can significantly kill liver cancer cells, melanoma cells, pancreatic cancer cells and leukemia cells (Lam TL, Wong GK, Chong HC, Cheng PN, Choi SC, Chow TL, et al. Recombinant human Arginase...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/50A61K45/06A61K45/00A61P35/00
Inventor 林辰鞠佃文赵舒薇李力
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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