A functionalized dendrimer-encapsulated gold nanoparticle/pd-l1 siRNA complex and its preparation and application

A technology of dendrimers and amine dendrimers, which is applied in the field of multifunctional polymer nanocarriers and their preparation and application, to achieve good application potential, mild experimental conditions, and good gene transfection ability

Active Publication Date: 2022-07-12
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Retrieval of related literature and patents at home and abroad shows that siPD-L1 is loaded with fluorescamine-modified PEGylated polyamide-amine dendrimer-coated gold nanoparticles as a carrier for immunotherapy of tumors, but has not been reported yet.

Method used

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  • A functionalized dendrimer-encapsulated gold nanoparticle/pd-l1 siRNA complex and its preparation and application
  • A functionalized dendrimer-encapsulated gold nanoparticle/pd-l1 siRNA complex and its preparation and application
  • A functionalized dendrimer-encapsulated gold nanoparticle/pd-l1 siRNA complex and its preparation and application

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Experimental program
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Effect test

Embodiment 1

[0049] (1) Weigh 50 mg of the fifth-generation polyamidoamine dendrimer (PAMAM) and dissolve it in 50 mL of ultrapure water. Weigh M-SCM-2000 (78 mg) and dissolve it in 7.8 mL of ultrapure water. Ultrasonic for 15 minutes Then quickly added to G5.NH 2 In the solution, the reaction was stirred at room temperature for 4 hours to obtain a crude G5-PEG product.

[0050] (2) 687 μL HAuCl was added dropwise to the G5-PEG obtained above 4 (30mg / mL), after stirring and mixing in an ice bath for 20 minutes, excess pre-cooled NaBH was added 4 The solution (5.67 mg, 1 mg / mL) was reacted for 4 hours to obtain {G5-PEG-(Au 0 ) 25 }crude product.

[0051] (3) The {G5-PEG-(Au 0 ) 25 } Dissolve in pH 8 phosphate buffer (25 mL), then dropwise add 680 μL of fluorescamine solution (4 mg / mL, dissolved in methanol), react in a dark, 40°C water bath and stir for 30 minutes, then use the interception A dialysis bag with a molecular weight of 5000 was dialyzed against the aqueous solution for 3...

Embodiment 2

[0053] Take 6 mg of {G5-PEG-(Au 0 ) 25 -F} dissolve in 600 μL of D 2 O, the solution was sonicated for 10 min, and then the H NMR spectrum of the material was measured using an NMR spectrometer. NMR results such as figure 2 shown, the proton peak at chemical shift 2.25-3.4 ppm represents G5.NH 2 The methylene proton peak of 3.63ppm is (M-SCM-2000) repeating unit -CH 2 CH 2 - The proton peaks of each G5.NH are calculated from the ratio of their integrated areas 2 There are 16.8 M-SCM-2000 molecules connected to it, which is close to its feeding molar ratio G5.NH 2 : M-SCM-2000 (1:20). The proton peak at 7.2-7.6ppm is attributed to the proton on the benzene ring of fluorescein, and the proton peak at 8.63ppm is attributed to the amino group (-NH) of fluorescein and the dendrimer. 2 ) protons on -COOH generated after the reaction, according to the ratio of their integral areas, calculate each G5.NH 2 There are 3.8 fluorescamine molecules connected to it, which is close ...

Embodiment 3

[0055] The {G5-PEG-(Au 0 ) 25 } Dissolved in ultrapure water, prepared into a solution with a concentration of 200 μg / mL, and detected the UV absorption value in the range of 300nm-800nm ​​by UV-Vis spectrophotometry. UV-Vis spectrophotometric detection results are as follows image 3 As shown, {G5-PEG-(Au 0 ) 25 } There is an absorption peak around 520 nm, indicating the successful encapsulation of gold nanoparticles.

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Abstract

The invention relates to a functionalized dendrimer-encapsulated gold nanoparticle / PD-L1 siRNA complex and its preparation and application. The complex is a polyamide-amine dendrimer loaded with polyethylene glycol and fluorescamine on the surface and coated with gold nanoparticles on the surface to load PD-L1 siRNA. The method includes: preparation of G5-PEG solution, preparation of fluorescein-modified PEGylated dendrimer-encapsulated gold nanoparticles, and preparation of functionalized dendrimer-encapsulated gold nanoparticles / PD-L1 siRNA complex. The complex can effectively silence the expression of PD-L1 and realize tumor immunotherapy, which has good application potential in tumor immunotherapy. The method is simple, the experimental conditions are mild, and the operation is easy.

Description

technical field [0001] The invention belongs to the multifunctional macromolecular nanocarrier and the field of preparation and application thereof, and particularly relates to a functionalized dendrimer-encapsulated gold nanoparticle / PD-L1 siRNA complex and a preparation method and application thereof. Background technique [0002] Cancer is one of the main factors threatening human health. Traditional treatment methods include chemotherapy, radiotherapy and surgery, but there are adverse reactions such as tumor metastasis and side effects. In immunotherapy, immune checkpoint blockade therapy has become a popular tumor treatment method at this stage due to its good efficacy and long-term inhibition of tumor metastasis. One of the immune checkpoints with more in-depth research on the mechanism of action is PD-1 / PD-L1. When PD-1 expressed on the surface of T cells binds with PD-L1 expressed in tumor tissue, it will transmit negative effects to T cells. To regulate the signal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/713A61P35/00A61K49/00A61K33/242A61K47/34A61K9/51
CPCA61K31/713A61P35/00A61K49/0021A61K49/0054A61K33/242A61K9/5146A61K2300/00Y02A50/30
Inventor 史向阳薛雪李锦
Owner DONGHUA UNIV
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