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Kidney-targeted drug-loaded exosome, application and medicine for treating kidney diseases

A technology for kidney disease and exosomes, applied in the field of biomedicine, can solve problems such as poor targeting, achieve improved treatment effects, good application prospects, and reduce drug toxicity and side effects

Inactive Publication Date: 2021-12-28
姜海涛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poorly targeted treatments for kidney cancer and other kidney-related diseases

Method used

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  • Kidney-targeted drug-loaded exosome, application and medicine for treating kidney diseases
  • Kidney-targeted drug-loaded exosome, application and medicine for treating kidney diseases
  • Kidney-targeted drug-loaded exosome, application and medicine for treating kidney diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The kidney-targeted drug-loaded exosomes are obtained by introducing drugs for treating kidney diseases into kidney-tissue-targeted exosomes, and the kidney-tissue-targeted exosomes are derived from bladder cells.

[0029] Immortalized human bladder epithelial cells SV-HUC-1 were collected (the immortalized human bladder epithelial cells SV-HUC-1 were taken as an example in this example). SV-HUC-1 cell culture medium is cultured with exosome-free serum and DMEM medium; centrifuge at 2000g centrifugal force for 10min, take supernatant; then centrifuge at 10000g centrifugal force for 30min, take supernatant, discard cell debris and sediment; the centrifuged cell culture solution was centrifuged at 100,000g for 2 h, the precipitate was collected by resuspending in sterile PBS, and stored at 4°C for a short period of time to obtain kidney-targeted exosomes; extracted and separated Kidney-targeted exosome surface marker proteins and particle size distribution such as figure...

Embodiment 2

[0031] The exosomes secreted by SV-HUC-1 cells were taken and stained with PKH67 to trace the distribution of exosomes secreted by SV-HUC-1 cells in vivo. The specific steps are as follows:

[0032] Take 100 μg of exosomes secreted by SV-HUC-1 cells, incubate with 1 μL of PKH67 overnight at 4 °C in the dark, centrifuge at 100,000 g for 2 h, discard the supernatant, wash twice with PBS, no The exosomes secreted by SV-HUC-1 cells were resuspended in PBS and injected into C57bl / 6 mice through the tail vein (in this example, male C57bl / 6 (4-6 weeks) mice were purchased from Beijing Huafukang Biotechnology Co., Ltd., all mice were bred in SPF facilities); after 24 hours, the mice were anesthetized, and the hearts, livers, spleens, lungs, kidneys, stomachs and intestines of the mice were taken to make frozen sections, stained with Hoechst33342 After the cell nucleus, the biodistribution of exosomes secreted by SV-HUC-1 cells in various organs of the mouse was observed; the results s...

Embodiment 3

[0034] In this example, microRNA 1271 mimic, a drug used to treat kidney cancer, was selected.

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

[0036] Step 1: Mix 150 μg of kidney-targeted exosomes with microRNA 1271 mimic, make up to 150 μL with electroporation buffer (PBS, DMEM, Cytomix, Tris-HCl), and transfer to different sizes (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 kidney-targeted exosomes and microRNA 1271 mimic respectively;

[0037] Step 2: The electroporation product was ultracentrifuged at 100,000 g for 120 min, and the supernatant was collected to measure the drug loading.

[0038] Results: For microRNA 1271 mimic, 150 μg kidney-targeted exosomes were electroporated with microRNA 1271 mimic at a...

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Abstract

The invention belongs to the technical field of biological medicine, and discloses a kidney-targeted drug-loaded exosome, application and a medicine for treating kidney diseases. The kidney-targeted drug-loaded exosome is obtained by introducing the medicine for treating the kidney diseases into an exosome with kidney tissue targeting property. The exosome with kidney tissue targeting property is derived from bladder cells. The exosome secreted by the bladder cells can be enriched in the kidney without any modification, and the exosome secreted by the bladder cells solves the problem of exosome yield and has a good application prospect; and the exosome secreted by the bladder cells can be used for loading different medicines or active molecules, and through targeted kidney tissue administration, the treatment effect on the kidney 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, and in particular relates to a kidney-targeted drug-loaded exosome and a drug for the application and treatment of kidney diseases. Background technique [0002] Kidney disease is a general term for common diseases that seriously endanger human health, mainly including different types of nephritis, acute renal failure, kidney stones, renal cysts and so on. Kidney tumors are one of the most common tumors in the human urinary and male reproductive system. The general treatment principle for renal cell carcinoma is to perform surgical treatment for early stage patients without metastases or only locally advanced patients, and no surgery for advanced metastatic renal cell carcinoma. Opportunity, should adopt comprehensive treatment based on internal medicine. In terms of medical treatment, the use of targeted therapy for renal cell carcinoma has greatly prolonged the overall survival. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K47/46A61K45/00A61P13/12
CPCC12N5/0685A61K47/46A61K45/00A61P13/12
Inventor 姜海涛王天怡
Owner 姜海涛
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