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A kind of amphotericin b albumin nano-preparation and its preparation method and application

A technology of nano-albumin and amphotericin, which is applied in the field of medicine, can solve the problems of toxicity, unstable liposome manufacturing cost, and easy leakage of drugs, and achieve simple preparation process steps, minimize drug activity damage, and avoid toxicity. side effects

Active Publication Date: 2021-09-03
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, factors such as the easy leakage of drugs in liposome formulations, the instability of liposomes, and the high cost of their manufacture greatly limit the wide application of
[0005] On the other hand, the preparation method of adding other groups will bring unpredictable toxicity. For example, in the prior art such as CN104490847A, the method of introducing and adding other groups is used to realize the reconstruction and cross-linking of the internal disulfide bond of albumin. Amphotericin B albumin nano-formulations prepared in this way have a greater safety risk
In addition, the traditional preparation of albumin carrier does not make full use of the unique interaction between albumin carrier and drug, such as CN109771656A, making its drug loading effect often unsatisfactory

Method used

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  • A kind of amphotericin b albumin nano-preparation and its preparation method and application
  • A kind of amphotericin b albumin nano-preparation and its preparation method and application
  • A kind of amphotericin b albumin nano-preparation and its preparation method and application

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Effect test

Embodiment 1

[0037] Dissolve 40 mg of bovine serum albumin, 20 mg of sodium dodecyl sulfate (SDS) and 1.48 mg of dithiothreitol (DTT) in 1 mL of ultrapure water and stir at 90°C for 2 hours to form a heat-denatured reduction albumin. Albumin preparation process see figure 1 , dilute the concentration of the reduced albumin solution from 40 mg / mL to 1 mg / mL with 1 mL of 0.1 M MES buffer (pH=4.8), add 25 μL of 4 mg / mL amphotericin B (AmB) dissolved in DMSO, at 37 Stir at ℃ for 4.5 hours. Dialysis was performed with a dialysis bag with a molecular weight cut-off of 8000-14000 in deionized water at 25° C. for 48 hours, followed by freeze-drying to obtain amphotericin B albumin nanoparticle freeze-dried powder. figure 2 a shows the hydrated particle size distribution of amphotericin B albumin nanoparticles (AmB-NP). As can be seen from the figure, the average particle size was 59 nm, and the polydispersity coefficient (PDI) was 0.272.

Embodiment 2

[0039] 120 mg of bovine serum albumin, 55 mg of sodium dodecyl sulfate (SDS) and 4.5 mg of dithiothreitol (DTT) were dissolved in 3 mL of ultrapure water and stirred at 90°C for 2 hours to form reduced albumin. The concentration of the reduced albumin solution was diluted from 40 mg / mL to 1 mg / mL with 1 mL of 0.1 M MES buffer (pH=4.25), 20 μL of 4 mg / mL amphotericin B (AmB) dissolved in DMSO was added, and the solution was heated at 37°C. Stir under conditions for 4 hours. Dialysis was performed with a dialysis bag with a molecular weight cut-off of 8000-14000 in deionized water at 20° C. for 48 hours, followed by freeze-drying to obtain amphotericin B albumin nanoparticle freeze-dried powder. figure 2b shows the hydrated particle size distribution of amphotericin B albumin nanoparticles (AmB-NP). The average particle size was 41 nm and the polydispersity coefficient (PDI) was 0.281.

Embodiment 3

[0041] 50 mg of bovine serum albumin, 40 mg of sodium dodecyl sulfate (SDS) and 3 mg of dithiothreitol (DTT) were dissolved in 1 mL of ultrapure water and stirred at 95°C for 4.5 hours to form reduced albumin. The concentration of the reduced albumin solution was diluted from 40 mg / mL to 1 mg / mL with 1 mL of 0.1 M MES buffer (pH=5.0), 10 μL of 1 mg / mL amphotericin B (AmB) dissolved in DMSO was added, and the solution was heated at 45°C. Stir under conditions for 2 hours. Dialysis was performed with a dialysis bag with a molecular weight cut-off of 8000-14000 in deionized water at 25° C. for 48 hours, followed by freeze-drying to obtain amphotericin B albumin nanoparticle freeze-dried powder. The average particle size was 35 nm and the polydispersity coefficient (PDI) was 0.267.

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Abstract

The present invention relates to a kind of amphotericin B albumin nano-preparation and its preparation method and application, wherein through the effect of heating, reducing agent and surfactant, destroy the hydrophobic area of ​​protein and intramolecular disulfide bond, form free thiol-rich Albumin molecules form albumin nanoparticles through the intermolecular disulfide bond network and hydrophobic interaction. In the process of forming the intermolecular disulfide bond network, the small molecule antibacterial drug amphotericin B is introduced to form Protein Nanoformulations. Compared with the prior art, the present invention makes full use of the high binding force between amphotericin B and albumin, transforms its existing deficiencies into advantages, and is expected to exert great application potential in anti-infection therapy, through in vitro and In vivo experiments showed that the nanosystem altered the biodistribution of amphotericin B and reduced drug accumulation in the kidney.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to an amphotericin B albumin nano preparation and a preparation method and application thereof. Background technique [0002] Serum albumin has the advantages of safety, non-toxicity, good biocompatibility, low immunogenicity, and biodegradability, and its excellent solubility can improve the solubility of hydrophobic drugs (Chen Q, et al., 2016). Albumin Paclitaxel Nanoparticle Injection Suspension Approved by the US FDA in 2005, albumin, a new drug carrier, has attracted more and more researchers' attention. It has been proved to have good therapeutic effect in the treatment of non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer and pancreatic cancer. [0003] The preparation of albumin nanoparticles mainly includes the following methods: desolvation method, emulsification method, spray drying method, pH coagulation method, thermal gelation method, etc....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/51A61K47/42A61K31/7048A61P31/10
CPCA61K9/5169A61K31/7048A61P31/10
Inventor 李永勇安毛毛韩毅慎慧陈思敏
Owner TONGJI UNIV
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