a g5-mos 2 Preparation method of /bcl-2 siRNA complex

A technology of bcl-2siRNA and g5-mos2, which is applied in the field of preparation of G5-MoS2/Bcl-2siRNA complex, achieves the effects of easy operation, easy synthesis and purification, good dispersion and biocompatibility

Active Publication Date: 2020-05-29
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Retrieval of relevant literature and patents at home and abroad shows that the method of using molybdenum disulfide modified by the fifth-generation polyamidoamine dendrimers as a carrier to achieve dual therapy of photothermal and gene therapy has not been reported yet.

Method used

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  • a g5-mos <sub>2</sub> Preparation method of /bcl-2 siRNA complex
  • a g5-mos <sub>2</sub> Preparation method of /bcl-2 siRNA complex
  • a g5-mos <sub>2</sub> Preparation method of /bcl-2 siRNA complex

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Embodiment 1

[0054] (1) Weigh 100mg (NH 4 ) 2 MoS 4 The powder was dissolved in 20 mL of deionized water, stirred magnetically for 20 min, and then ultrasonically oscillated for 10 min until the powder was completely dissolved in water. While stirring, add 0.454mL of (N 2 h 4 )·H 2 O, ultrasonic vibration for 30min. The resulting mixture was poured into a 50mL polytetrafluoroethylene-lined reactor, then heated to 200°C for 10 hours, and the reactor was taken out to cool naturally to room temperature to obtain a black solution. Centrifuge the above black solution at 10,000 rpm for 5 minutes to collect the product, which is the obtained MoS 2 . MoS was washed with water and centrifuged 2 Purification was carried out, and after repeated 10 times, the purified MoS 2 Dissolve in 10mL deionized water and store at 4°C for later use.

[0055] (2) 3.996mg LA was dissolved in 5mL DMSO, then 51.81mg EDC·HCl and 33.18mg NHS were weighed, dissolved in 2mL DMSO solution, and stirred for 3h to ...

Embodiment 2

[0059] Carry out NMR characterization to the G5-LA prepared in embodiment 1 step (2), 1 HNMR characterization results such as figure 1 As shown, there is a proton peak at the chemical shift of 1.5-2.5ppm, which is a characteristic group proton peak in the molecular structure of LA. According to its and G5.NH 2 The integral area ratio between each G5.NH can be calculated 2 11.4 LA molecules were attached to it. The UV-vis results are shown in Figure 2, it can be seen that the modified G5.NH2 Before and after, the absorption characteristics of the ultraviolet absorption peak did not change significantly, indicating that the G5.NH 2 modification did not change the MoS 2 light absorption properties. TEM results such as image 3 As shown, the prepared MoS 2 (3a) presents layered structure, and modified G5.NH 2 After (3b), G5-MoS 2 A polymer is formed, the shape is close to spherical, and the particle size of the formed nanoflower is about 100-200nm. FESEM results such as ...

Embodiment 3

[0061] The MoS prepared by the method of step (1) and step (3) of embodiment 1 2 and G5-MoS 2 Dilute with PBS to obtain a solution with a concentration of 0.1 mg / mL, and take 1 mL of the solution for hydrodynamic diameter (6a) and surface potential (6b) characterization by a Malvern laser particle size analyzer (Malvern, MK, 633nm laser). The result is as Figure 6 As shown, the unmodified MoS 2 With large hydrodynamic diameter and negative surface potential, it is not suitable for gene transfection. while G5-MoS 2 The particle size of G5.NH was significantly reduced, indicating that 2 The modification improves the dispersion of the material. Meanwhile, G5-MoS 2 The charge carried on the surface has also changed from negative to positive, making it easier to be phagocytized by cells.

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Abstract

The invention relates to a preparation method of a G5-MoS2 / Bcl-2 siRNA compound. The preparation method comprises the steps that LA is activated by EDC.HCl and NHS to obtain activated LA; then, the activated LA is dropwise added to a G5.NH2 solution, reaction is performed for 24 hours, and dialysis and freeze-drying are performed to obtain G5-LA; the G5-LA is added to a MoS2 solution, ultrasonic oscillation and stirring are performed for 12 hours, and centrifugation and washing are performed to obtain G5-MoS2 nanoflowers; the G5-MoS2 nanoflowers and Bcl-2 SiRNA are incubated for 20-430 minutes to obtain the G5-MoS2 / Bcl-2 siRNA compound. The method has the advantages of being simple in process, easy to operate, high in photothermal conversion efficiency, simple in transfection condition and high in transfection efficiency and the like and has the good application prospect in photothermal and gene therapy.

Description

technical field [0001] The invention belongs to the field of preparation of nanomaterials in photothermal and gene dual therapy, in particular to a kind of G5-MoS 2 Preparation method of / Bcl-2 siRNA complex. Background technique [0002] The exploration of cancer treatment methods is a process of continuous development and challenges. Traditional treatment methods such as surgery, chemotherapy and radiotherapy have many shortcomings, which are reflected in the following aspects: first, the tumor site cannot be completely removed, which may lead to tumor regeneration and metastasis; second, long-term Chemotherapy may lead to multi-drug resistance of tumors; third, there is also damage to normal cells, which can cause side effects such as nausea, hair loss and fatigue. Therefore, it is very important to develop a new type of tumor treatment system. Such a system needs to be friendly to the organism, and at the same time have high tumor treatment efficiency and low side eff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K41/00A61K47/60A61P35/00
CPCA61K41/0052A61K48/0041A61K48/0091
Inventor 史向阳孔令丹孙文杰
Owner DONGHUA UNIV
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