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Lung-targeted drug-loaded exosome, application and medicine for treating lung diseases

A lung disease and exosome technology, applied in the field of biomedicine, can solve the problems of lack of targeting, poor patient tolerance, high toxicity and side effects, and achieve the goal of improving therapeutic effect, good application prospects, and reducing drug toxicity Effect

Inactive Publication Date: 2021-10-08
姜海涛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, whether it is chemical drugs or biomolecules, cancer treatment involves the following three problems. First, due to the lack of targeting, the drugs have a lethal effect on normal tissues, resulting in high toxic side effects and poor patient tolerance.
Second: Because many drug molecules are water-soluble, it is difficult to be taken up by cells, and it is difficult to reach an effective drug concentration in the cell; Third: The stability of drug molecules in the body, before reaching the lesion, there will be many enzymes in the body It is degraded, the stability of biological macromolecules is poor, and it is easy to be degraded in the body
The expression of survivin was detected in early embryonic cells, indicating that survivin is involved in the development of normal embryonic tissues, however, when the tissue matures, it is difficult to detect the expression of survivin, and there is almost no expression in most adult tissues

Method used

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  • Lung-targeted drug-loaded exosome, application and medicine for treating lung diseases
  • Lung-targeted drug-loaded exosome, application and medicine for treating lung diseases
  • Lung-targeted drug-loaded exosome, application and medicine for treating lung diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Collect normal human colorectal tissue epithelial cells FHC (this embodiment takes normal human colorectal tissue epithelial cell FHC as an example). Take the normal human colorectal tissue epithelial cell FHC culture medium (colorectal cell culture medium is cultured with exosome-free serum and DMEM medium) and centrifuge at 2000g centrifugal force for 10min, take the supernatant; then centrifuge at 10000g centrifugal force After 30 minutes, take the supernatant, discard the cell debris and sediment; centrifuge the centrifuged cell culture solution at 100,000 g (g is the acceleration of gravity) for 2 h, resuspend the precipitate with sterile PBS, and store it at 4°C for a short period of time. Obtain lung-targeted exosomes. The extracted and isolated lung-targeted exosome surface marker protein and particle size distribution are as follows: figure 1 , figure 2 shown. The expression of exosomes secreted by colorectal cells was measured, and the concentration of exo...

Embodiment 2

[0030] Exosomes secreted by FHC from normal human colorectal tissue epithelial cells were collected and stained with PKH67 to track the distribution of exosomes secreted by colorectal cells in vivo. The specific steps are as follows:

[0031] Take 100 μg of exosomes secreted by colorectal cells, incubate with 1 μL PKH67 overnight at 4°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 colorectal cells were resuspended and injected into C57bl / 6 mice through the tail vein. It should be noted that: in this example, male C57bl / 6 (4-6 weeks) mice (4-6 weeks ) was purchased from Beijing Huafukang Biotechnology Co., Ltd. All mice were bred in SPF facilities; 24 hours later, the mice were anesthetized, and the hearts, livers, spleens, lungs, kidneys, stomachs and intestines of the mice were taken for frozen sections , after staining the nucleus with Hoechst...

Embodiment 3

[0033] In this example, the drug siRNA-survivin used to treat lung diseases is selected as an example, and the sequence of siRNA-survivin is: 5'-GAAUUUGAGGAAACUGCGATT-3'

[0034] 5'-UCGCAGUUUCCUCAAAUUCTT-3' was synthesized in Shanghai Gemma Company.

[0035] In this example, drugs for treating lung diseases are introduced into lung-targeted exosomes to prepare drugs for lung tumor-targeted therapy. The specific operations are as follows:

[0036]1) Take 150 μg of lung-targeted exosomes and mix them with fluorescent siRNA-survivin. The electrotransfer buffer can be (PBS, DMEM, Cytomix, Tris-HCl) to make up to 150 μL, and transfer to different specifications (0.2cm, 0.4cm) in the electric cup. Different waveforms (exponential wave, square wave) and different voltages (50V, 100V, 200V, 300V) were used to electroporate lung-targeted exosomes and siRNA-survivin respectively;

[0037] 2) The electroporation product was ultracentrifuged for 120 min under a centrifugal force of 100,...

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Abstract

The invention belongs to the technical field of biological medicine, and discloses a lung-targeted drug-loaded exosome, application and a medicine for treating lung diseases; the medicine for treating the lung diseases is introduced into the exosome with lung tissue targeting property to obtain the lung-targeted drug-loaded exosome; the exosome with the lung tissue targeting property is derived from colorectal cells; and a lung tumour treatment medicine contains the lung-targeted drug-loaded exosome. Compared with the prior art, the exosome secreted by the colorectal cells can be enriched in the lung without any modification; the exosome derived from the colorectal cells is used; the problem of exosome yield is solved; the application prospect is very good; the exosome from the colorectal cells can be used for loading different medicines or active molecules; and through targeted lung tissue administration, the treatment effect on lung diseases is improved; and toxic and side effects of the medicines are reduced.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a lung-targeted drug-loaded exosome and a drug for the application and treatment of lung diseases. Background technique [0002] The lungs are one of the most important organs in the human body, and abnormalities of the lungs can cause various lung diseases. Including bronchitis, pulmonary fibrosis, emphysema, pulmonary abscess, pulmonary embolism, chronic obstructive pulmonary disease, pneumonia, tuberculosis, pneumothorax, bullae, pulmonary nodules, lung tumors, etc. Lung cancer is the most common primary malignant tumor of the lung. The vast majority of lung cancers originate from the bronchial mucosal epithelium, so it is also called bronchial lung cancer. Over the past 50 years, the morbidity and mortality of lung cancer have risen rapidly in countries all over the world, especially in industrialized countries, and lung cancer has ranked first among male patients who di...

Claims

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Application Information

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IPC IPC(8): A61K47/46A61K45/00A61K9/50A61P11/00C12N5/071
CPCA61K47/46A61K45/00A61P11/00A61K9/5068C12N5/0679
Inventor 姜海涛王天怡
Owner 姜海涛
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