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Novel composite nano preparation based on sonodynamic/immune combined therapy and preparation method and application thereof

A nano-preparation and combination therapy technology, applied in nanotechnology, nanotechnology, nano-drugs, etc., can solve the problems of poor water solubility, lack of tumor targeting in vivo distribution, etc.

Active Publication Date: 2021-06-11
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the poor water solubility of chlorin e6 and its derivatives and the lack of tumor targeting in vivo distribution, nanosystems are used to enhance the drug’s targeted delivery ability, increase its accumulation in the tumor area, and achieve sustained drug release. is an effective means

Method used

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  • Novel composite nano preparation based on sonodynamic/immune combined therapy and preparation method and application thereof
  • Novel composite nano preparation based on sonodynamic/immune combined therapy and preparation method and application thereof
  • Novel composite nano preparation based on sonodynamic/immune combined therapy and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Comparative group 1: Preparation of liposome product Ⅰ

[0033] (1) Accurately weigh 20mg of soybean lecithin, 5mg of DSPE-PEG5k, and sound sensitizer Ce6Me 3 Dissolve 1mg in a 50ml round-bottomed flask with 5ml chloroform until completely dissolved, then evaporate under reduced pressure on a rotary evaporator to form a film, and dry it overnight in a vacuum oven.

[0034](2) Add 4 ml of PBS for hydration, and stir in a water bath at 50° C. for 1.5 h with a magnetic stirrer (400 rpm) under nitrogen protection.

[0035] (3) Pass the resulting solution through polycarbonate membranes of 0.45 μm and 0.22 μm through an extruder in a water bath at 50° C. to obtain liposome product I: liposome-encapsulated sound sensitizer (Lip-Ce6Me 3 ).

[0036] Comparative group 2: Preparation of liposome product II

[0037] (1) Accurately weigh 20mg of soybean lecithin, 5mg of DSPE-mPEG5k, and sound sensitizer Ce6Me 3 1mg and 1.5mg of cinnamaldehyde derivatives were dissolved in a 5...

Embodiment 2

[0045] Carry out basic characterization to the nano liposome that embodiment 1 makes

[0046] (1) Performance test experiment 1: Liposome Lip-Ce6Me 3 - Morphological analysis of CA-MPLA

[0047] Liposome Lip-Ce6Me was obtained by transmission electron microscope (TEM, HT7700EXALENS) 3 - TEM image of CA-MPLA as figure 2 Shown in (a). Take 1mL liposome solution Lip-Ce6Me 3 -CA-MPLA, the hydrated particle size measured by Zeta potential and particle size analyzer (Malvern Zetasizer, Nanozs90) is 149.1nm, and the polydispersity coefficient (Poly DiversityIndex, "PDI") is 0.168, and the particle size distribution diagram is as follows figure 2 Shown in (b). Lip-Ce6Me was measured by the same method 3 -MPLA and Lip-Ce6Me 3 -The particle size change graph of CA-MPLA within 15 days, such as figure 2 As shown in (c), it shows that the nanoliposome is relatively stable within 15 days.

[0048] (2) Performance test experiment 2: Liposome Lip-Ce6Me 3 -CA-MPLA absorption spect...

Embodiment 3

[0053] (1) MTT test nanoliposome in vitro cytotoxicity and in vitro anti-tumor efficacy

[0054] In order to evaluate the cytotoxicity in vitro, breast cancer cells 4T1 were cultured in a 96-well plate, and 4T1 cells in the logarithmic growth phase were taken, and 8×10 3 Cells were incubated overnight in a 37°C incubator, and different concentration gradients of Lip-Ce6Me3 and Lip-Ce6Me were added 3 -CA(Ce6Me 3 : 10μg / mL, 30μg / mL, 50μg / mL, 100μg / mL, 200μg / mL;), compare the ultrasound toxicity and non-ultrasound toxicity, namely: Lip-Ce6Me 3 ,Lip-Ce6Me 3 (+US), Lip-Ce6Me 3 -CA, Lip-Ce6Me 3 -CA(+US). After adding the drug, incubate for 4 hours, wash off the drug with pre-cooled PBS, and re-add the complete medium, for Lip-Ce6Me 3 (+US), Lip-Ce6Me 3 -CA(+US) group was subjected to ultrasonic treatment, and the ultrasonic intensity was 3W / cm 2 , 10min. Put the 96-well plate into the cell culture incubator and incubate for 24 hours, discard the culture medium in the 96-wel...

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Abstract

The invention discloses a novel composite nano preparation based on sonodynamic / immune combined therapy and a preparation method and application thereof, the novel composite nano preparation comprises a sound-sensitive agent, an immune activator, an active oxygen enhancing drug and liposome, the sound-sensitive agent and the active oxygen enhancing drug are entrapped in a liposome phospholipid bimolecular layer, and the immune activator is embedded on the lipidosome phospholipid bimolecular layer. The method comprises the following steps: (1) preparing a sound-sensitive agent, a cinnamyl aldehyde derivative, MPLA, a lecithin and DSPE-PEG5k into a solution; (2) evaporating out the solvent in the solution obtained in the step (1), and dispersing the drug-loaded lipid film in a buffer solution to obtain a drug-loaded liposome suspension; and (3) enabling the solution obtained in the step (2) to pass through a polycarbonate film to obtain the composite nano-liposome. The preparation can induce anti-tumor reaction, not only can prevent the development of in-situ solid tumors, but also can prevent the tumors from metastasis to far-end tissues. The composition is especially suitable for a combined treatment preparation of sonodynamic therapy and immunotherapy.

Description

technical field [0001] The invention belongs to the technical field of multifunctional nanomaterials, and in particular relates to a preparation method of a novel composite nano-preparation combined with immunotherapy and sonodynamic therapy and its anti-tumor application. Background technique [0002] Cancer poses a serious threat to human life and health. According to the World Cancer Report 2020 of the World Health Organization (WHO), in the next two decades, the number of cancer cases worldwide may increase by 60%. In low- and middle-income countries, the increase may be as high as 81%, and effective treatment of cancer is urgently needed. The current clinical treatment methods for cancer mainly include surgery, chemotherapy, and radiotherapy. However, the above-mentioned traditional therapies not only have limited therapeutic effects, but also tend to impose a great burden on the body. Once the tumor metastasizes, it is even more difficult to cure. [0003] Sonodynami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K41/00A61K31/11A61K39/39A61K9/127A61K47/24A61P35/00B82Y5/00
CPCA61K41/0033A61K31/11A61K31/357A61K39/39A61K9/1271A61K47/24A61P35/00B82Y5/00A61K2039/55572A61K2039/585A61K2300/00
Inventor 邵堃上官晓燕李广哲赵伟杰王阳王柳张羽萱
Owner DALIAN UNIV OF TECH
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