A nanomaterial-based autophagy blocking system and its preparation method, and its application in the treatment of solid tumors with arsenic drugs

A technology of nanomaterials and solid tumors, applied in drug combinations, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve problems such as unsatisfactory therapeutic effects of arsenic drugs

Active Publication Date: 2021-05-14
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a nanomaterial-based autophagy blocking system and its preparation method, as well as its application in the treatment of solid tumors with arsenic drugs, so as to solve the unsatisfactory therapeutic effect of arsenic drugs on solid tumors in the prior art The problem

Method used

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  • A nanomaterial-based autophagy blocking system and its preparation method, and its application in the treatment of solid tumors with arsenic drugs
  • A nanomaterial-based autophagy blocking system and its preparation method, and its application in the treatment of solid tumors with arsenic drugs
  • A nanomaterial-based autophagy blocking system and its preparation method, and its application in the treatment of solid tumors with arsenic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Comparison of the efficacy of ATO on acute promyelocytic leukemia cells NB4 and human liver cancer cells HepG2

[0047] ATO was purchased from Sigma (product number: 202673), dissolved in 0.1M NaOH solution, adjusted to pH 8 with 1M HCL, and sterilized by filtration.

[0048] Human hepatoma cells HepG2 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in 1640 medium (Gibco) containing 10% fetal bovine serum. 37°C, 5% CO 2 , cultured in saturated humidity. cells in 7×10 4 / well density inoculated in 24-well plates and adhered overnight. ATO was added at a concentration of 2, 4, 8, 16, and 32 μM, and cells without any treatment were used as controls. Each group had three replicate wells. After incubation for 48 hours, MTT (Sigma) was stained for 4 hours, and 10% acidic SDS was added to dissolve the generated cells. Formazan was crystallized, and the UV absorbance value of each well was measured at OD 570nm, and the c...

Embodiment 2

[0051] Example 2: Evaluation of curative effect of ATO combined with autophagy blocking drug CQ on tumor cells.

[0052] CQ was purchased from Sigma (product number: C6628) and dissolved in sterile water.

[0053] In Western blot experiments, HepG2 cells were 7 × 10 4 Inoculate at a density of 24 wells per well and adhere to the wall overnight. Set up the following experimental groups, each with three replicate wells: CQ (12.5 μg / mL), ATO (2 μM and 4 μM each), CQ+ATO mixture (12.5 μg / mL CQ+2 μM ATO and 12.5 μg / mL CQ+ 4μM ATO for each group), and the cells without any treatment were used as the control, and incubated for 48h. Wash twice with PBS, lyse the cells with 1×SDS loading buffer, denature at 95°C, transfer to PVDF membrane after polyacrylamide gel electrophoresis. The PVDF membrane was blocked with 6% skimmed milk powder dissolved in PBST (0.1% Tween 20) for 1 h, washed 3 times with PBST, and a 1:1000 primary antibody was added. The primary antibodies involved are a...

Embodiment 3

[0056] Example 3: The autophagy regulation effect of nanodiamond on cells.

[0057] Nano-diamonds (NDs, Gansu Jinshi Nano Materials Co., Ltd.), the purity of the nano-diamonds is >99%, the diameter of a single particle is about 2-10 nm, and a cluster structure of about 250 nm is formed in the solution. UV irradiation for 30min, dispersed in sterile water.

[0058] In Western blot experiments, HepG2 cells were 7 × 10 4 Inoculate in a 24-well plate at a density of 24 wells, and adhere to the wall overnight. 50 μg / mL NDs and 12.5 μg / mL CQ were added respectively, and the cells without any treatment were used as the control. Each group had three replicate wells and incubated for 48 hours. Cell lysis, protein electrophoresis, and membrane transfer methods are the same as in Example 2. The primary antibodies involved in this example are as follows: anti-LC3 (Novus), P62 (Abcam) and GAPDH (Abcam), mTOR (cell signaling technology), P-mTOR (cell signaling technology), p70S6K (cell s...

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Abstract

The invention provides a nanomaterial-based autophagy blocking system, a preparation method thereof, and an application in arsenic drug treatment of solid tumors. The preparation method comprises the following steps: 1) dissolving the arsenic trioxide in NaOH solution and adjusting the pH to 8-8.5 to prepare the arsenic trioxide solution; 2) dispersing the nanomaterial in water, physiological saline or a buffer system to prepare the nanomaterial suspension solution; and 3) mixing the arsenic trioxide solution with the nanomaterial suspension to prepare an autophagy blocking system. The autophagy blocking system based on nanomaterials provided by the present invention can effectively block the autophagy of solid tumor cells induced by ATO, thereby significantly improving its killing effect on solid tumor cells, so that ATO can produce solid tumors at low concentrations. Good therapeutic effect, less toxic and side effects on normal tissues. The invention provides a method capable of significantly improving the therapeutic effect of ATO in solid tumors, and has good medical application prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a nanomaterial-based autophagy blocking system and a preparation method thereof, as well as an application in treating solid tumors with arsenic drugs. Background technique [0002] Arsenic trioxide (ATO) is a traditional Chinese medicine in my country. Professors Chen Zhu and Wang Zhenyi used this drug to treat acute promyelocytic leukemia (APL). They conquered APL for the first time. The clinical results showed that the cure rate of patients was as high as 95%. In addition to APL cells, ATO also has a certain killing effect on a variety of solid tumor cells, but its effect is far less than that of APL cells. Autophagy is one of the common responses of solid tumor cells against chemotherapy. Cells in a state of autophagy will reduce the activity of related enzymes required to promote cell death, thereby greatly reducing the effect of chemotherapy. ATO can induce au...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K33/36A61K33/44A61K33/242A61K33/26A61P35/00
CPCA61K33/24A61K33/26A61K33/36A61K33/44A61K2300/00
Inventor 樊春海诸颖崔之芬
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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