Anti-tumor synergic medicine

An anti-tumor and drug technology, applied in the field of anti-tumor drugs, can solve problems that have not yet been seen

Inactive Publication Date: 2014-08-20
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] So far, there have been no reports on the induction of tumor cell autophagy by recombinant human a

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Preparation of recombinant human arginase

[0028] The coding sequence encoding human arginase is obtained by using the PCR method, and the coding sequence of the gene is 969 bp in full length, with enzyme cutting 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, extract the recombinant plasmid, linearize the recombinant vector with BglⅡ enzyme digestion, transform Pichia pastoris by electric shock, pick positive transformants, and screen through RDB, MM and MD plates to obtain the phenotype: The clone of his+muts was further induced to express on the shaker level, and the expression of arginase on the shaker level was detected b...

Embodiment 2

[0032] Example 2 Preparation of autophagy inhibitor drugs

[0033] (1) Preparation of chloroquine: Dissolve an appropriate amount of chloroquine in pure water to make a 10mmol / L stock solution, filter and sterilize with a 0.1μm filter and store at 4°C. Dilute 500-1000 times with PRMI-1640 medium for in vitro experiments Used to inhibit autophagy;

[0034] (2) Preparation of ammonium chloride: Dissolve an appropriate amount of ammonium chloride in water to make a 0.4mol / L storage solution, filter and sterilize with a 0.1μm filter, and store at 4°C. In vitro experiments were diluted 50-80 times to inhibit cell autophagy;

[0035] (3) Preparation of hydroxychloroquine: Dissolve an appropriate amount of hydroxychloroquine in pure water to make a 10mmol / L storage solution, filter and sterilize with a 0.1μm filter, store at 4°C, and dilute 500-1000 times in the in vitro laboratory to inhibit autophagy;

[0036] (4) Preparation of 3-MA: Take an appropriate amount of 3-MA dry pow...

Embodiment 3

[0040] Example 3: Cell Culture

[0041] Raji and Daudi lymphoma cells were cultured in PRMI-1640 medium containing 10% FBS and 1% penicillin-streptomycin. When the cells grew to the logarithmic growth phase, the cells were centrifuged at 1000 rpm, the supernatant was discarded, and fresh Culture medium Gently blow the cells evenly, with 2*10 5 The concentration of cells / ml was transferred to the cell culture plate for culture, and the cells in each group were given the corresponding concentration of recombinant human arginase treatment; when used, 2mmol / L of 3-MA and 5mmol / L of CQ were added 1h before administration. Inhibit the autophagy of Raji and Daudi cells, and proceed to the next step after continuous culture for 24h, 48h or 72h.

[0042]

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Abstract

The invention relates to anti-tumor medicines, and concretely relates to an anti-tumor synergic medicine. The synergic medicine is a medicinal complex consisting of one or a plurality of cell autophagy inhibitors and recombined human argininase. The cell autophagy inhibitor(s) and the recombined human argininase are administrated in a combined administration manner or in a sequential administration manner. By inhibiting autophagy of tumor cells induced by argininase, the antagonistic action, caused by cell autophagy, of tumor on argininase treatment is offset, and therefore the killing effect of recombined human argininase on tumor is enhanced, and the treatment effect of argininase on tumor is substantially enhanced. The synergic medicine is applicable to lymphoma, leukemia, liver cancer, lung cancer, melanoma, breast cancer and myeloma.

Description

technical field [0001] The present invention relates to anti-tumor drugs, in particular to an anti-tumor synergistic drug, in particular to a drug complex composed of an autophagy inhibitor and recombinant human arginase. Background technique [0002] The prior art discloses that most of the chemotherapeutic drugs currently used for tumor treatment will produce drug resistance and have strong side effects on patients. Studies have reported that biomacromolecular drugs developed in recent decades have the advantages of high targeting, high efficiency, and low toxicity and side effects. Among them, the anti-tumor effect of recombinant human arginase has been widely demonstrated, but its in vivo affinity Low, the effective dose is relatively large, and there are still some potential toxic and side effects. [0003] Arginase is a key enzyme involved in the urea cycle in the liver of mammals. In the urea cycle, arginine is converted into ornithine and urea. Studies have shown t...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K38/50A61P35/00A61P35/02
Inventor 鞠佃文曾贤李玉彬范佳君王子玉王绍飞
Owner FUDAN UNIV
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