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Mhs cell-derived exosome-loaded forsythin drug delivery system and its application

A drug delivery system and exosome technology, applied in the field of biomedicine, can solve the problems of poor absorption and low bioavailability, and achieve the effect of prolonging the half-life, good anti-tumor metastasis effect, and good anti-tumor cell metastasis effect

Active Publication Date: 2021-11-02
HENAN UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, Phil has poor oral absorption and low bioavailability

Method used

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  • Mhs cell-derived exosome-loaded forsythin drug delivery system and its application
  • Mhs cell-derived exosome-loaded forsythin drug delivery system and its application
  • Mhs cell-derived exosome-loaded forsythin drug delivery system and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Preparation of MHS cell-derived exosomes loaded with forsythin drug delivery system (Phil-Exos)

[0022] 1. Reagents and cells

[0023] Fetal bovine serum (FBS), American Gibco Company; RPMI 1640 medium, Hyclon Company; Forsythin reference substance, batch number MUST-14052103, mass fraction ≥ 98%, Chengdu Institute of Biology, Chinese Academy of Sciences; pre-stained protein Marker (10- 180kDa), American Thermo Company; SDS-PAGE gel preparation kit, blocking solution BSA, Kangwei Century Company; CD63, Alix, β-Actin (internal reference) antibody, HRP-labeled IgG, Wuhan Sanying Biotechnology Co., Ltd.; PBS (pH=7.4), penicillin-streptomycin (double antibody, 100IU·mL -1 ), RIPA cell lysate, BCA protein quantification kit, ECL Plus ultra-sensitive luminescence solution, Beijing Solarbio Technology Co., Ltd.; PKH67 staining kit, Beyotime; DAPI staining solution, DiI staining kit, Beyotime Company; 4% poly Formaldehyde general-purpose tissue fixative, biosharp ...

Embodiment 2

[0030] Particle size and Zeta potential measurement of embodiment 2, MHS-Exos and Phil-Exos

[0031] Take 100 μL of MHS-Exos and Phil-Exos suspensions respectively, dilute them to 1 mL with ultrapure water, and equilibrate at room temperature for 2 minutes, then measure their particle size and Zeta potential with a Malvern nanoanalyzer, and repeat 3 times.

[0032] see results figure 1 , the average particle size of MHS-Exos is (126.78±1.98)nm, the dispersion coefficient of PDI is 0.332±0.061, the Zeta potential is (-18.25±0.05)mV; the average particle size of Phil-Exos is (138.93±0.98)nm, the PDI The dispersion coefficient is 0.237±0.023; the Zeta potential is (-19.33±0.17)mV. The particle size results showed that exosomes had a narrow particle size distribution range, the particle size increased slightly after drug loading, and the potential did not change significantly before and after drug loading. The operation of drug loading had little effect on the surface protein con...

Embodiment 3

[0033] Embodiment 3, transmission electron microscope observation form

[0034] Take 10 μL of the MHS-Exos and Phil-Exos suspension and drop it on the copper grid, let it stand for 1 min, then blot the remaining liquid with filter paper, then add 3 drops of 3% phosphotungstic acid solution for negative staining at room temperature for 5 min, and blot the remaining liquid with the filter paper. Dry at room temperature and observe under a transmission electron microscope, and take pictures to record the shape of exosomes.

[0035] see results figure 2 , the diameter of exosomes slightly increased after loading Phil, which was consistent with the results of Malvern particle size. Both MHS-Exos and Phil-Exos showed a round or oval shape with a complete membrane structure, ranging in size, with a diameter of about 30-150nm. The particles have good dispersibility, and the background is clean and clear.

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Abstract

The invention relates to the technical field of biomedicine, in particular to a MHS cell-derived exosome-loaded forsythin drug delivery system and its application. The drug delivery system is prepared by mixing MHS cell-derived exosomes with forsythin solution, incubation, ice bath ultrasound, constant temperature culture recovery, centrifugation, washing and resuspension, centrifugation, and resuspension; The concentration of cystin was 400-410 μg / mL; in terms of protein amount, the concentration of MHS cell-derived exosomes was 400-410 μg / mL. The Phil-Exos prepared by the invention is well taken up by A549 cells, has a high inhibitory effect on the migration of A549 cells, and significantly improves the anti-tumor cell migration ability of Phil.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an MHS cell-derived exosome loading forsythin drug delivery system and its application. Background technique [0002] Lung cancer is a malignant tumor of the lung with the highest morbidity and mortality. Forsythia is the dry fruit of Forsythiasuspense (Thunb.) Vahl, a plant of Oleaceae. It is a lignin component of forsythia, which has various pharmacological activities such as antibacterial and anti-oxidation, anti-lung cancer, anti-virus, liver protection, and anti-inflammation, and has application and development prospects. Zheng Mo et al found that Phil can inhibit the development of lung tumors by down-regulating the expression of vascular endothelial growth factor and endostatin in lung cancer tissues. However, Phil has poor oral absorption and low bioavailability. Therefore, how to improve the stability and bioavailability of Phil and further improve the effect of P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/50A61K31/7048A61K47/46A61P35/00A61P35/04
CPCA61K9/5068A61K31/7048A61P35/00A61P35/04
Inventor 黄海英王磊吕田田张晓艳余海艳薛丙权申骐菲
Owner HENAN UNIV OF CHINESE MEDICINE
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