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Gene delivery system capable of demixing cell membrane coating and its preparation method and application

A technology of gene delivery and cell membrane, which is applied in gene therapy, medical preparations of non-active ingredients, capsule delivery, etc., can solve the problems of nanocomposite internalized drug release obstacles, etc., to prolong blood circulation time, promote interaction, The effect of improving serum stability

Active Publication Date: 2022-04-15
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this cell membrane coating technology also has certain problems, that is, after accumulation at the lesion site, the membrane itself will also become an obstacle for the internalization of the nanocomplex by the target cells and the release of the drug due to the negative charge on the membrane surface.

Method used

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  • Gene delivery system capable of demixing cell membrane coating and its preparation method and application
  • Gene delivery system capable of demixing cell membrane coating and its preparation method and application
  • Gene delivery system capable of demixing cell membrane coating and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Prepare the cationic polypeptide material, as follows:

[0058] First, the ring-opening polymerization of five-membered ring N-carboxyanhydride (NCA) was initiated by hexamethyldisilane (HMDS), and a polypeptide (polyγ-propargyl-L- Glutamic acid, PPLG), and the molecular weight of PPLG was characterized by gel chromatography (GPC) ( figure 1 ). Then in the glove box, PPLG (20 mg, 0.06 mmol), guanidinium small molecule azide N 3 -HG (85 mol%, 0.204 mmol), small molecule azide compound containing benzene ring (15 mol%, 0.036 mmol), pentamethyldiethylenetriamine (PMDETA, 126 μL, 0.604 mmol) were dissolved in dimethyl Formamide (DMF, 4 mL), then copper bromide (CuBr, 0.12 mmol, 17.56 mg) was added. After stirring the reaction at room temperature for 48 h, hydrochloric acid solution (1 M, 1 mL) was added, dialyzed in deionized water for 3 days (MWCO = 3.5 kDa), and the product was obtained as a white solid after freeze-drying. 400 M H NMR spectrum ( 1 H NMR) to characte...

Embodiment 2

[0068] Polypeptide P-Ben (1 mg / mL) and siRNA (0.1 μg / mL) were dissolved in DEPC water, mixed according to the mass ratio of P-Ben / siRNA to 15, and incubated at room temperature for 30 min to form a binary nanocomposite Object P-Ben / siRNA (BS). PLL-CA (2 mg / mL) was dissolved in DEPC water (pH = 8, adjusted with sodium hydroxide), added to the BS complex solution according to the mass ratio of PLL-CA / P-Ben as 2, 5, and 10, and mixed well , incubated at room temperature for 30 min to form a ternary nanocomplex PLL-CA / P-Ben / siRNA (BSPC), with Figure 6 structure shown.

Embodiment 3

[0070] The mixed membrane solution (HM) of Example 1 was added to the BSPC complex solution of Example 2 according to the mass ratio of cell membrane / PLL-CA of 0.2, and then ultrasonicated for 3 min (ultrasonic frequency 40 kHz) to obtain detachable mixed cell membrane packets The gene delivery system (BSPC@HM), with Figure 7 In the structure shown, the mass ratio of cell membrane, polymer material, cationic polypeptide and drug is 15:75:15:1.

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Abstract

The invention provides a gene delivery system capable of demixing cell membrane coating, its preparation method and application. The system uses helical polypeptide material to load gene drug (Sav1 siRNA) to form a binary nanocomposite core, then wraps a layer of polymer material to form a ternary nanocomposite, and finally coats a platelet-macrophage mixed cell membrane. Form the final nanocomposite system. The system has damage site targeting and serum stability, and under weakly acidic conditions, the outer layer of the coated cell membrane is shed, exposing the internal binary nanocomplex, increasing the cell uptake by enhancing the interaction with the cell membrane, and realizing Efficient transfection of target genes. This system successfully realizes the mixed coating of two kinds of cell membranes and the process of specific membrane removal at the damaged site, and has great application prospects in the field of gene delivery.

Description

technical field [0001] The invention relates to the field of gene loading and delivery, in particular to the construction and application of a demixable cell membrane-coated gene delivery system, which is used for siRNA transfection. Background technique [0002] Gene vectors are important tools for loading nucleic acid molecules, delivering them into target cells and successfully expressing them. Cationic polymers are one of the important categories of non-viral gene carriers, which can achieve efficient transmembrane delivery of gene drugs. However, the surface of the cationic polymer is positively charged, which will adsorb negatively charged proteins in the blood and cause the aggregation of nanocomplexes, so it cannot be applied to systemic drug delivery. In recent years, nanocomposites coated with biological cell membranes not only retain the physicochemical properties of the synthesized nanoparticle cores, but also possess the unique biological functions of the encap...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K9/51A61K47/46B82Y5/00
CPCA61K48/0025A61K9/5176B82Y5/00
Inventor 殷黎晨梁秋君
Owner SUZHOU UNIV
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