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Albumin nanoparticles capable of double-stage oxygen carrying and preparation method and application thereof

A technology of albumin nanoparticles and albumin, which is applied in the field of radiotherapy sensitization drugs to achieve high-efficiency oxygen delivery, reduce systemic side effects of drugs, and improve radiotherapy sensitivity

Active Publication Date: 2019-05-28
南京从一医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there is no report on the application of synergistic perfluorocarbons and catalase to improve tumor hypoxia and achieve radiosensitization

Method used

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  • Albumin nanoparticles capable of double-stage oxygen carrying and preparation method and application thereof
  • Albumin nanoparticles capable of double-stage oxygen carrying and preparation method and application thereof
  • Albumin nanoparticles capable of double-stage oxygen carrying and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of perfluorocarbon-catalase composite albumin nanoparticles

[0026] Measure 1mL of human serum albumin stock solution (10g / 50mL) into a vial, add deionized water to dilute to 1.9mL, and mix well. Add 300 μL of perfluorotripentylamine (that is, the volume ratio of perfluorotripentylamine to the total preparation system is 10%) and 100 μL of ethanol (analytical grade), and mix well by pipetting. Place the nanoemulsion obtained above in an ice-water bath, use an ultrasonic probe to penetrate into the vial to near the bottom of the liquid, the ultrasonic power is 300W, the duty cycle is 50%, and the duration is 2 minutes. After the ultrasonic is finished, place the preparation in an ice-water bath Let cool for 1-2 minutes. Add 700 μL of catalase solution with a mass concentration of 30 mg / mL into the above system (that is, the proportion of catalase in the total preparation system is 7 mg / mL), and continue to operate according to the above ultrasoni...

Embodiment 2

[0031] Example 2: Investigation of the Oxygen Carrying and Slow Oxygen Release Ability of Perfluorocarbon-Calase Composite Albumin Nanoparticles

[0032] Take 200 μL of the perfluorocarbon-catalase composite albumin nanoparticle solution in Example 1, dilute it to 3 mL with deionized water, and take another 3 mL of deionized water as a control. The two groups were placed in the small chamber and filled with pure oxygen for 20s, and then the oxygen content in the solution and the oxygen content of each solution after 30min and 24h were detected in real time with an oxygen electrode.

[0033] The results show that after oxygenation, the oxygen content in pure water is 356 μmol / L, and the oxygen content of the perfluorocarbon-catalase composite albumin nanoparticle solution is 606 μmol / L, which shows that the prepared nanoparticle solution It has excellent oxygen carrying capacity. After 30 minutes, the oxygen content of pure water and nanoparticle solution changed to 293 μmol / L...

Embodiment 3

[0034] Example 3: Oxygen generation catalyzed by perfluorocarbon-catalase composite albumin nanoparticles in hydrogen peroxide solution

[0035] Catalase detection kit was used to detect hydrogen peroxide catalytic activity of perfluorocarbon-catalase composite albumin nanoparticles. Take 20 μL of the sample into a 1.5ml centrifuge tube (use free catalase solution as a positive control), add catalase detection buffer to 40 μL, and mix well. Add 10 μL of 250 mM hydrogen peroxide solution and mix quickly with a pipette. After reacting at 25°C for 1-5 minutes, add 450 μL catalase reaction termination solution, take another centrifuge tube and add 40 μL catalase detection buffer, then add 10 μL of the above reaction system that has been terminated and mixed, and mix well. Add 10 μL to a 96-well plate, then add 200 μL of chromogenic working solution, incubate at 25°C for at least 15 minutes, and measure the absorbance at 520 nm of the solution.

[0036] The results show that (see ...

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Abstract

The invention belongs to the field of radiotherapy sensibilization medicines, and relates to albumin nanoparticles capable of double-stage oxygen carrying and a preparation method and application thereof. The albumin nanoparticles are mainly prepared from albumin, perfluoro-carbon and catalase, the particle size of the albumin nanoparticles is even, the property is stable, the biosecurity is good,and the problem of radiotherapy resisting of solid tumors can be solved. Experimental results show that the nanoparticles can target the tumors and improve tumor hypoxia by means of double-stage oxygen carrying, for specific performance of double-stage oxygen carrying of the nanoparticles, the high-oxygen-carrying perfluoro-carbon directly supplies oxygen (first stage oxygen carrying) to the hypoxic tumors directly through oxygen concentration gradient difference, meanwhile, the catalase conducts catalytic hydrolysis on high-concentration hydrogen peroxide on the tumor portions to generate oxygen for indirect oxygen carrying (second stage oxygen carrying), through synergism of first stage oxygen carrying and second stage oxygen carrying, a tumor hypoxia microenvironment is reversed, and effective sensibilization is conducted on oxygen-dependent tumor radiotherapy.

Description

technical field [0001] The invention relates to an albumin nanoparticle capable of two-stage oxygen delivery, a preparation method and application thereof, and belongs to the field of radiotherapy sensitization drugs. Background technique [0002] Clinical studies have shown that hypoxia occurs in more than 90% of solid tumors. This is because the malignant tumor grows rapidly and grows disorderly, so the demand for oxygen is greatly increased. At the same time, the angiogenesis of the tumor is insufficient, and the blood supply in the tumor tissue is relatively insufficient, so most solid tumors have hypoxic areas. [0003] Radiation therapy, as one of the three main means of tumor treatment, is widely used in tumor treatment. Oxygen plays an important biological effect in killing tumor cells by ionizing radiation. Oxygen is a sensitizer for cells sensitive to radiation. Water generates hydrated electrons and free radicals under the action of radiation. Free radicals can d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K51/08A61K51/12A61K38/44A61K33/16A61P35/00
Inventor 吴锦慧张宝丽王浩然陈维生
Owner 南京从一医药科技有限公司
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