Colorectum-targeted drug-loaded exosome and application thereof, and drug for treating colorectal diseases

A colorectal and exosome technology, applied in the field of biomedicine, can solve problems such as poor targeting, toxic and side effects of chemotherapy drugs, and achieve the effects of reducing toxic and side effects of drugs, improving therapeutic effects, and promising application prospects.

Inactive Publication Date: 2021-10-22
姜海涛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Chemotherapy drugs have good anti-tumor activity, but due to their poor targeting, chemotherapy drugs have extremely toxic side effects

Method used

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  • Colorectum-targeted drug-loaded exosome and application thereof, and drug for treating colorectal diseases
  • Colorectum-targeted drug-loaded exosome and application thereof, and drug for treating colorectal diseases
  • Colorectum-targeted drug-loaded exosome and application thereof, and drug for treating colorectal diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] An embodiment of the present invention provides a colorectal-targeted drug-loaded exosome, which is introduced into the colorectal tissue-targeted exosome by a drug for treating colorectal diseases The exosomes with colorectal tissue targeting were obtained from lung cells.

[0030] Human embryonic lung fibroblasts HFL-1 were collected (in this example, human embryonic lung fibroblasts HFL-1 were used as an example).

[0031] It should be noted that the HFL-1 cell culture medium is cultured with exosome-free serum and DMEM medium; centrifuge at 2000g centrifugal force for 10min, take the supernatant; then centrifuge at 10000g centrifugal force for 30min, take the supernatant, discard Cell debris and sediment; the centrifuged cell culture medium was centrifuged under the centrifugal force of 100000g (g is the acceleration of gravity) for 2 hours, the precipitate was collected by resuspending in sterile PBS, and stored at 4°C for a short period of time to obtain colorecta...

Embodiment 2

[0033] The exosomes secreted by HFL-1 cells in the above-mentioned Example 1-2 were stained with PKH67 to track the distribution of exosomes secreted by HFL-1 cells in the body. Male C57bl / 6 mice (4-6 Zhou) were purchased from Beijing Huafukang Biotechnology Co., Ltd., and all mice were bred in SPF facilities. Specific steps are as follows:

[0034] Take 100 μg of exosomes secreted by HFL-1 cells, incubate overnight with 1 μL KH at 674°C in the dark, centrifuge at 100,000 g (g is the acceleration of gravity) for 2 h, discard the supernatant, wash twice with PBS, and then wash with sterile PBS Exosomes secreted by lung cells were resuspended and injected into C57bl / 6 mice via tail vein. After 24 hours, the mice were anesthetized, and the hearts, liver, spleen, lungs, kidneys, stomach and intestines of the mice were taken to make frozen sections. After the nuclei were stained with Hoechst33342, the biological activity of exosomes secreted by HFL-1 cells in each organ of the mic...

Embodiment 3

[0036]The drugs for treating colorectal diseases include but not limited to siRNA, microRNA, protein, antibody, fluorouracil, oxaliplatin, irinotecan, capecitabine. In this example, the drug oxaliplatin for the treatment of colorectal cancer was selected.

[0037] Importing drugs for treating colorectal diseases into colorectal-targeted exosomes to prepare colorectal tumor-targeted therapeutic drugs, the specific operations are as follows:

[0038] Take 150 μg of colorectal-targeted exosomes and mix them with oxaliplatin. The electrotransfer buffer can be PBS, DMEM, Cytomix, Tris-HCl. In this example, PBS is used to make up to 150 μL, and transferred to different specifications (0.2 cm, 0.4cm) in the electric transfer cup. Different waveforms (exponential wave, square wave) and different voltages (50V, 100V, 200V, 300V) were used to electroporate colorectal-targeted exosomes and oxaliplatin;

[0039] The electroporation product was ultracentrifuged at 100,000 g for 120 min, ...

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Abstract

The invention belongs to the technical field of biological medicine, and provides a colorectum-targeted drug-loaded exosome and application thereof, and a medicine for treating colorectal diseases. The colorectum-targeted drug-loaded exosome is obtained by introducing the medicine for treating the colorectal diseases into an exosome with a colorectal tissue targeting property; and the exosome with colorectal tissue targeting property is derived from lung cells. A colorectal tumor treatment drug contains the colorectum-targeted drug-loading exosome. Compared with the prior art, the exosome secreted by the lung cells can be enriched in the colorectum without any modification, the exosome derived from the lung cells is used, the problem of exosome yield is solved, and good application prospect is obtained; and different drugs or active molecules can be loaded by using the lung cell-derived exosome, and the treatment effect of colorectal diseases is improved and toxic and side effects of the drugs are reduced through targeted colorectal tissue administration.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a colorectal-targeted drug-loaded exosome and a drug for its application and treatment of colorectal diseases. Background technique [0002] The colon and rectum are parts of the digestive system that make up a long, muscular tube called the large intestine. Cancers that start in the colon are called colon cancers, and cancers that start in the rectum are called rectal cancers. Both can also be called colorectal cancer. Colorectal cancer is a common malignant tumor in the gastrointestinal tract and is the fourth largest tumor type in the world, causing 500,000 deaths per year. The incidence and mortality of colorectal cancer in my country are 27.3 / 100,000 and 13.9 / 100,000, respectively, ranking fifth in the incidence and mortality of tumors, and patients over 45 years old account for 93.5% of the total affected population. The early symptoms are not obvious, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K47/46
CPCC12N5/0688A61K47/46
Inventor 姜海涛王天怡
Owner 姜海涛
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