Preparation method of copper porphyrin-folate liposome nanoparticles and application thereof as sound-sensitive agent

A nanoparticle and folic acid technology, applied in the field of biomedicine, can solve the problems of poor water solubility of metalloporphyrin small molecular compounds, affecting the research of sonosensitizers, and being easily metabolized, etc., to improve water solubility and tumor targeting, improve Biofilm penetration, improve the effect of enrichment

Pending Publication Date: 2021-03-09
GUANGDONG MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, metalloporphyrin small molecule compounds have problems such as poor water so

Method used

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  • Preparation method of copper porphyrin-folate liposome nanoparticles and application thereof as sound-sensitive agent
  • Preparation method of copper porphyrin-folate liposome nanoparticles and application thereof as sound-sensitive agent
  • Preparation method of copper porphyrin-folate liposome nanoparticles and application thereof as sound-sensitive agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of folate-targeted copper porphyrin liposome nanoparticles (FA-CuPP):

[0039] ① Weigh 5 mg of copper porphyrin and dissolve it in 1 mL of analytically pure methanol to obtain a copper porphyrin methanol solution with a concentration of 5 mg / ml; ② Weigh 20 mg of soybean lecithin and dissolve it in 5 mL of analytically pure chloroform; ③ Weigh out PEGylated 2 mg of folic acid liposome (DSPE-PEG-folate) was dissolved in 5 mL of analytically pure chloroform; ③ the methanol solution of copper porphyrin and the folic acid liposome solution were added to the soybean lecithin solution; ④ rotary evaporation removed the organic Solvent, to obtain a uniform lipid film, vacuum drying for 24 hours, remove the remaining solvent; ⑤ add 10mL of ultrapure water, ultrasonic hydration in ultrasonic cleaning wave for 10 minutes, to obtain liposome suspension; ⑥ use ultrasonic breaker for intermittent Sonicate for 10 minutes; the copper porphyrin-folate liposome nanoparticles (...

Embodiment 2

[0041]Preparation of folic acid-targeted liposome-loaded copper porphyrin nanoparticles (FA-CuPP): Weigh 1 mg of copper porphyrin and dissolve in 500 μL of analytically pure methanol; ② Weigh 10 mg of soybean lecithin and PEGylated folic acid lipid Dissolve 1 mg of DSPE-PEG-folate in 1 mL of analytically pure chloroform; ③ After mixing the above solutions, remove the organic solvent by rotary evaporation to obtain a uniform lipid film, and dry it in vacuum for 24 hours to remove the residual organic solvent; ④Add 5mL of ultrapure water, and ultrasonically hydrate for 5 minutes in an ultrasonic cleaning wave to obtain a nanoparticle suspension; ⑤Use an ultrasonic breaker to perform intermittent ultrasonication for 5 minutes; Protect from light after filtering with a filter membrane for later use.

[0042] The experiments showed that soy lecithin and PEGylated folate liposomes formulated alone or together had no effect on the results of folate-targeted copper porphyrin liposome ...

Embodiment 3

[0046] Example 3 Application of Copper Porphyrin-Folic Acid Liposome Nanoparticles as Sonosensitizer.

[0047] (1) Targeting detection of copper porphyrin-folate liposome nanoparticles in tumors: at 37°C, 1×10 5 The 4T1 tumor cells per well were seeded in 6-well plates, and after incubation overnight, nanometers containing FA-CuPP (FA-CuPP prepared in Example 1) and L-CuPP (L-CuPP prepared in Comparative Example 1) were replaced respectively. After incubation for 3 hours in the culture medium of the particles (the concentration of CuPP is 10 μg / mL), the cells were washed three times with phosphate buffer, and the cells were collected to detect the difference of phagocytosis by flow cytometry. Depend on Figure 4 It can be seen that the uptake of folic acid-modified copper porphyrin liposome nanoparticles in tumors is significantly stronger than that of non-folate-modified copper porphyrin liposome nanoparticles, indicating that FA-CuPP has good tumor targeting.

[0048] (2) ...

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Abstract

The invention discloses a preparation method of copper porphyrin-folate liposome nanoparticles and application thereof as a sound-sensitive agent. The preparation method comprises the following steps:dissolving copper porphyrin with methanol, dissolving lecithin and folate liposome in chloroform, mixing the two solutions, preparing a lipid film by a rotary evaporation method, and carrying out ultrasonic hydration with ultrapure water to synthesize the copper porphyrin-folate liposome nanoparticles. The nanoparticles have excellent targeting property in tumor cells with high expression of folate receptors, and are beneficial to enrichment at tumor sites so as to improve the anti-tumor effect; and under ultrasonic excitation, the copper porphyrin absorbs sound energy to generate transitionand converts surrounding oxygen into singlet oxygen to kill tumor cells. The invention provides the sound-sensitive agent capable of generating singlet oxygen through ultrasonic excitation to kill thetumor cells, and the folate targeted liposome is used as a carrier to carry the sound-sensitive agent, so that the water solubility and the targeting property are improved, and the sonodynamic therapy (SDT) effect is further improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine. Specifically, it is a preparation method of copper porphyrin-folate liposome nanoparticles and its application as a sound sensitizer. Background technique [0002] Sonodynamic therapy (SDT) is to excite the sound sensitizer by ultrasound to generate reactive oxygen species such as singlet oxygen or free radicals to kill tumor cells and achieve the purpose of inhibiting tumor growth. Because ultrasound has a strong penetration depth and can penetrate body tissues and focus precisely on the lesion, sonodynamic therapy is considered to be a very promising non-invasive treatment technology, and has become a research hotspot in the field of tumor treatment in recent years. Since sonodynamic therapy is an emerging treatment technology, related research is still very limited, and sound sensitizers are the key to the development of sonodynamic therapy. At present, the commonly used sonosensitizers ...

Claims

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Application Information

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IPC IPC(8): A61K41/00A61K9/127A61K47/24A61K47/22A61P35/00
CPCA61K41/0033A61K9/1271A61K47/24A61K47/22A61K9/1277A61P35/00
Inventor 马爱青陈华清刘建强闫冲尹婷郑明彬张丽姗冉慧
Owner GUANGDONG MEDICAL UNIV
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