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Preparation and application of milk exosome loaded icariin nano preparation

A technology of icariin and nano preparations, applied in nanotechnology, nanotechnology, nanomedicine, etc., can solve the problems of toxicity and high cost, achieve low toxicity, low cost, and improve efficacy and safety

Pending Publication Date: 2021-05-14
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The primary consideration for exosome-loaded drugs is the selection of donor cells for exosomes. Now, exosomes derived from immature dendritic cells and mesenchymal stem cells are usually used, but the amount of exosomes collected from the above sources is relatively large. Less, high cost, and certain toxicity (exosomes derived from mesenchymal stem cells)

Method used

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  • Preparation and application of milk exosome loaded icariin nano preparation
  • Preparation and application of milk exosome loaded icariin nano preparation
  • Preparation and application of milk exosome loaded icariin nano preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Extraction method of bovine milk exosomes

[0046] 1) Centrifuge fresh milk at 13000g in a 60mL centrifuge bottle for 30min at 4°C, and take the middle whey;

[0047] 2) 600-800 mesh vacuum pump filtration;

[0048] 3) Centrifuge at 100,000g for 2 hours at 4°C, and take the middle whey;

[0049] 4) 1000-2000 mesh vacuum pump filtration;

[0050] 5) Centrifuge at 13500g for 1.5h at 4°C, leave 1-2ml of the bottom, and collect in a 50ml EP tube;

[0051] 6) Centrifuge at 100,000 g for 2 hours at 4°C, discard the supernatant, and separate the milk exosome suspension;

[0052] 7) Mixing icariin and exosome suspension at a volume ratio of 1:9 at room temperature; freeze-drying to obtain the exosome-loaded icariin nanomedicine.

Embodiment 2

[0054] Identification of bovine milk exosomes

[0055] The size, structure and morphology of exosomes were observed by nanoparticle tracking analysis and transmission electron microscopy. The protein expression levels of exosome surface marker factors CD63, ALIX, CD81 and microcapsule surface marker CD40 were detected by Western Blot. Exosomes were labeled with PKH67 fluorescent staining, and the uptake of exosomes by MC3T3-E1 was detected.

Embodiment 3

[0057] Synthesis of Exosome-loaded Icariin Nanomedicine

[0058] Mix icariin and exosome suspension at a volume ratio of 1:9 at room temperature. After freeze-drying, the exosome-loaded icariin nanomedicine was obtained.

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Abstract

The invention provides an exosome loaded icariin nano preparation, which is prepared by loading icariin on cow's milk exosomes, and the preparation method comprises the following steps: performing centrifuging for 30-60 minutes by using a centrifugal machine at 4 DEG C and 13,000-15,000 g, taking middle layer whey, and performing filtering by using a vacuum pump with 600-800 meshes; performing centrifuging at the temperature of 4 DEG C at 100000-150000 g for 1-2 hours, and taking the whey in the middle layer; and performing filtering by using a 1000-2000 mesh vacuum pump, performing centrifuging for 1-2 hours at the temperature of 4 DEG C under the condition of 13000-15000g, reserving 1-2 ml of whey at the bottom, collecting the whey in a 50ml EP tube, performing centrifuging for 1-2 hours at the temperature of 4 DEG C under the condition of 100000-150000g, discarding the supernatant, and separating out the milk exosome; mixing icariin and the exosome suspension at room temperature according to a ratio of 1: 9; and performing freezing and drying to obtain the exosome loaded icariin nano-drug. The material has the unique advantages that the availability is high, the cost is low, the toxicity is low, the yield is high, the medicine can be orally delivered, the large-scale production can be achieved, the bioavailability of the oral medicine can be potentially enhanced, the efficacy and the safety of the medicine can be improved, and the bone repair capability of the hostoside can be improved.

Description

technical field [0001] The invention belongs to the technical field of pharmaceutical preparations, and more specifically relates to the preparation and application of a milk exosome-loaded icariin nano-preparation. Background technique [0002] Traditional Chinese medicine, as a traditional characteristic medicine in our country, has a good therapeutic effect on many diseases. Icariin, a natural antioxidant derived from traditional Chinese medicine, can improve the body's immunity and promote bone formation, and is widely used in the research of osteoporosis and bone defect-related diseases. [0003] However, due to the low solubility and low permeability of icariin, its bioavailability is low, and it is difficult to exert the best therapeutic effect. Clinically, it is necessary to develop a carrier that can improve its biological efficacy and facilitate the body's absorption, so as to help it fully exert its efficacy. Soluble in ethanol, ethyl acetate, insoluble in water...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61K9/50A61K47/46B82Y5/00A61P19/08
CPCA61K31/7048B82Y5/00A61K9/5068A61K9/0053A61P19/08
Inventor 牛卫东董明王丽娜白桦
Owner DALIAN MEDICAL UNIVERSITY
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