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Amphotericin B albumin nanometer preparation and preparation method and application thereof

A technology of albumin nanometer and amphotericin, which is applied in the field of medicine, can solve the problems of not making full use of the unique interaction force, unstable manufacturing cost of liposome, and unsatisfactory drug loading effect, so as to improve drug solubility, improve Toxic and side effects, the effect of avoiding toxic and side effects

Active Publication Date: 2020-08-14
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Amphotericin B albumin nanometer preparation and preparation method and application thereof
  • Amphotericin B albumin nanometer preparation and preparation method and application thereof
  • Amphotericin B albumin nanometer preparation and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Dissolve 40mg of bovine serum albumin, 20mg of sodium dodecyl sulfate (SDS) and 1.48mg of dithiothreitol (DTT) in 1mL of ultrapure water, and stir at 90°C for 2 hours to form a heat-denatured reduced 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 for 4.5 hours at °C. The amphotericin B albumin nanoparticle freeze-dried powder was obtained by dialyzing in deionized water at 25° C. for 48 hours with a dialysis bag with a molecular weight cut-off of 8000-14000, and then freeze-drying. figure 2 a shows the hydrated particle size distribution of amphotericin B albumin nanoparticles (AmB-NP). It can be seen from the figure that the average particle diameter is 59 nm, and the polydispersity index (PDI) is 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. 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=4.25), add 20 μL of 4 mg / mL amphotericin B (AmB) dissolved in DMSO, and incubate at 37 °C Stirred under conditions for 4 hours. The amphotericin B albumin nanoparticle freeze-dried powder was obtained by dialyzing in deionized water at 20° C. for 48 hours with a dialysis bag with a molecular weight cut-off of 8000-14000, and then freeze-drying. figure 2b shows the hydrated particle size distribution of amphotericin B albumin nanoparticles (AmB-NP). The average particle size is 41 nm, and the polydispersity index (PDI) is 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. 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=5.0), add 10 μL of 1 mg / mL amphotericin B (AmB) dissolved in DMSO, and incubate at 45 °C Stirred under conditions for 2 hours. The amphotericin B albumin nanoparticle freeze-dried powder was obtained by dialyzing in deionized water at 25° C. for 48 hours with a dialysis bag with a molecular weight cut-off of 8000-14000, and then freeze-drying. The average particle size is 35 nm, and the polydispersity index (PDI) is 0.267.

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Abstract

The invention relates to an amphotericin B albumin nanometer preparation and a preparation method and application thereof. Under the action of heating, a reducing agent and a surfactant, a hydrophobicregion and intramolecular disulfide bonds of proteins are destroyed, albumin molecules rich in free sulfhydryl are formed; through an intermolecular disulfide bond network and hydrophobic interaction, albumin nanometer particles are formed; in the process of forming the intermolecular disulfide bond network, micromolecule antibacterials amphotericin B is introduced, and the amphotericin B albuminnanometer preparation can be formed. Compared with the prior art, high binding force between amphotericin B and albumin is sufficiently used, defects existing in the prior art are converted into advantages, and the amphotericin B albumin nanometer preparation hopefully exerts great application potentials in anti-infective therapy. In vitro and in vivo experiments indicate that the nanometer system changes biological distribution of the amphotericin B, and accumulation of medicines in kidneys is reduced.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to an amphotericin B albumin nano-preparation and its preparation method and application. 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). Nab-paclitaxel Nanoparticle Injection Suspension It was approved by the FDA in 2005, which makes albumin, a new drug carrier, attract more and more researchers' attention. Studies have shown that it has good therapeutic effects 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,...

Claims

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Application Information

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Patent Type & Authority Applications(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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