Preparation method of anti-tumor microbubbles carrying chemotherapeutic drugs, medicament and application

A chemotherapy drug, anti-tumor technology, applied in the field of anti-tumor

Inactive Publication Date: 2020-10-23
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the anti-tumor vaccine prepared by this method can only prevent the recurrence of existing tumors, and is not effective in preventing multi-drug resistance of tumor cells.

Method used

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  • Preparation method of anti-tumor microbubbles carrying chemotherapeutic drugs, medicament and application
  • Preparation method of anti-tumor microbubbles carrying chemotherapeutic drugs, medicament and application
  • Preparation method of anti-tumor microbubbles carrying chemotherapeutic drugs, medicament and application

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preparation example Construction

[0035] The invention relates to a preparation method, medicament and application of anti-tumor microbubbles carrying chemotherapeutic drugs. The anti-tumor microbubbles are produced by treating tumor cells with radiation; the chemotherapeutic drugs are common clinical chemotherapeutic drugs. The anti-tumor microvesicles carrying chemotherapeutic drugs have broad-spectrum anti-tumor activity and can effectively kill tumor cells of autologous origin and non-autologous origin. It can effectively treat tumors that are not sensitive to chemotherapy or tumors that cannot be directly treated by radiotherapy, either locally or intravenously. Its anti-tumor principle mainly involves the combined tumor killing effect of a large amount of reactive oxygen species carried by microvesicles and chemotherapy drugs, as well as the regulation of the tumor immune microenvironment, such as promoting the reprogramming of macrophages. The anti-tumor microvesicles carrying chemotherapy drugs are com...

Embodiment 1

[0056] Embodiment 1, constructing and quantifying the microvesicle protein concentration after radiotherapy, and quantifying the dose of chemotherapeutic drugs carried by high performance liquid chromatography, and the preservation method of microbubbles carrying chemotherapeutic drugs after radiotherapy.

[0057] Cultivate tumor cells in a 10mm×10mm culture dish with 10% FBS (fetal bovine serum) medium. When the number of cells in the culture dish reaches about 5×106, perform radiotherapy at a dose of 20GY. On the first day after radiotherapy Change the medium, add 20ml medium containing 10% FBS and a certain concentration of chemotherapeutic drugs, collect all the liquid in the culture dish on the third day, and use gradient centrifugation to extract microvesicles after radiotherapy. Take the cell culture medium after radiotherapy at 1000g and centrifuge for 10min, then take the supernatant, then centrifuge the supernatant at 14000g for 2min to remove debris, discard the prec...

Embodiment 2

[0058] Example 2, the killing effect of microbubbles carrying chemotherapeutic drugs on tumor cells after radiotherapy.

[0059] Prepare 100 μl of cell suspension in a 96-well plate with a cell density of 5×104 cells / ml. The 96-well plate was pre-incubated for 24 hours in an incubator (37° C., 5% CO 2 ). Add different amounts of microbubbles (0.04, 0.08, 0.12, 0.16mg / ml) carrying chemotherapeutic drugs produced by radiotherapy to the culture plate. The plates were incubated in the incubator for 48 hours. 10 μl of CCK8 (Cell Counting Kit-8) solution was added to each well, and the culture plate was incubated in an incubator for 2 hours, and the absorbance at 450 nm was measured with a microplate reader. The absorbance value reflects the activity of the cells. Such as image 3 As shown, as the number of anti-tumor microbubbles carrying chemotherapy drugs increases after radiotherapy, the killing ability of the microbubbles on tumor cells is gradually enhanced, and the killin...

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Abstract

The invention relates to a preparation method of anti-tumor microbubbles carrying chemotherapeutic drugs, a medicament and application. The method comprises the following steps: 1) subjecting primarytumor cells to culture and passage; 2) carrying out X-ray irradiation on the primary tumor cells cultured in a culture dish obtained in the step 1), and continuing culture; and 3) co-incubating the cells obtained in the step 2) with chemotherapeutic drugs, and collecting supernatant to obtain the microbubbles carrying the chemotherapeutic drugs. The invention further provides the anti-tumor microbubbles carrying the chemotherapeutic drugs, and the anti-tumor microbubbles carrying the chemotherapeutic drugs are obtained by purifying and concentrating anti-tumor microbubbles. The anti-tumor microbubbles prepared by the preparation method of the anti-tumor microbubbles carrying the chemotherapy drugs can be used for preparing anti-tumor drugs, can achieve a more efficient killing effect on tumors, and expands the application range of radiotherapy and chemotherapy.

Description

Technical field: [0001] The present invention relates to the field of anti-tumor technology, in particular to a preparation method, medicament and application of anti-tumor microbubbles carrying chemotherapeutic drugs. Background technique: [0002] The bystander effect (radiation-induced bystander effect, RIBE) caused by radiation therapy refers to the death of not only the irradiated cells but also the surrounding cells that have not received the radiation after the cells are irradiated. The main principle of RIBE is that the microbubbles produced by irradiated cells contain high levels of ROS and some other DAMP damage molecules, and the microbubbles can be efficiently taken up by surrounding tumor cells, leading to indirect death of tumor cells. Previous studies have revealed that the microvesicles produced by injecting tumor cells alone after radiotherapy can effectively treat and delay the occurrence and development of tumors, which proves that autologous tumor cell mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/13A61K31/675A61K31/519A61K31/513A61K31/7068A61K31/704A61K31/407A61K33/243A61K9/127A61P35/00A61P35/04
CPCA61K9/1271A61K9/1277A61K31/407A61K31/513A61K31/519A61K31/675A61K31/704A61K31/7068A61K35/13A61P35/00A61P35/04A61K33/243A61K2300/00
Inventor 金红林
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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