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A folic acid-modified nano-preparation containing ccl19 recombinant expression gene and its preparation method and application

A technology for expressing genes and nano preparations, which is applied in the fields of folic acid-modified nano preparations containing CCL19 recombinantly expressed genes and the fields of their preparation and use, and achieves the effect of excellent application prospects.

Active Publication Date: 2022-05-24
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no report of a gene carrier capable of efficiently transfecting CCL19 into cells, and no report of an efficient antitumor drug containing the CCL19 gene

Method used

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  • A folic acid-modified nano-preparation containing ccl19 recombinant expression gene and its preparation method and application
  • A folic acid-modified nano-preparation containing ccl19 recombinant expression gene and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of folic acid-modified nanoformulations containing CCL19 recombinantly expressed gene

[0029] a. Dissolve 20mg DOTAP, 15mgFa-PEG-PLA and 65mg MPEG-PLA in 4ml acetone;

[0030] b. The organic solvent was removed under reduced pressure at 55°C for 1h, and then kept at room temperature and high vacuum for more than 6h;

[0031] c. Hydrate and remove the membrane with 5% glucose solution, the temperature of the water bath is 55°C, and the time is 1 h; after the membrane is removed, it is filtered with a 0.22 μm microporous membrane to obtain a nano-gene carrier, which is denoted as F-DMA;

[0032] d. F-DMA and pCCL19 (the gene sequence is shown in SEQ ID NO.1) were mixed in mass ratios of 25:1, 50:1, 100:1 and 150:1, and incubated at room temperature for 30 minutes to obtain a plasmid / nano combination substance, denoted as F-DMA / CCL19.

Embodiment 2

[0033] Example 2 Preparation of folic acid-modified nanoformulations containing CCL19 recombinantly expressed gene

[0034] a. Dissolve 10mg DOTAP, 15mgFa-PEG-PLA and 75mgMPEG-PLA in 4ml acetone;

[0035] b. The organic solvent was removed under reduced pressure at 55°C for 1 hour, and then kept at room temperature and high vacuum for more than 6 hours.

[0036] c. Hydrate and remove the membrane with 5% glucose solution, the temperature of the water bath is 55°C, and the time is 1 h; after the membrane is removed, it is filtered with a 0.22 μm microporous membrane to obtain a nano-gene carrier, which is denoted as F-DMA;

[0037] d. Mix F-DMA and pCCL19 at mass ratios of 25:1, 50:1, 100:1 and 150:1, and incubate at room temperature for 30 min to obtain a plasmid / nano composition, denoted as F-DMA / CCL19.

Embodiment 3

[0038] Example 3 Preparation of folic acid-modified nanoformulations containing CCL19 recombinantly expressed gene

[0039] a. Dissolve 15mg DOTAP, 70mgMPEG-PLA and 15mgFa-PEG-PLA in 5ml acetone;

[0040] b. The organic solvent was removed under reduced pressure at 55°C for 1 hour, and then kept at room temperature and high vacuum for more than 6 hours.

[0041] c. Hydrate and remove the membrane with 5% glucose solution, the temperature of the water bath is 55°C, and the time is 1 h; after the membrane is removed, it is filtered with a 0.22 μm microporous membrane to obtain a nano-gene carrier, which is denoted as F-DMA;

[0042] d. Mix F-DMA and pCCL19 at mass ratios of 25:1, 50:1, 100:1 and 150:1, and incubate at room temperature for 30 min to obtain a plasmid / nano composition, denoted as F-DMA / CCL19.

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Abstract

The invention discloses a folic acid-modified nano-preparation containing CCL19 recombinant expression gene, which is a nano-preparation formed by taking the CCL19 recombinant expression gene as an active ingredient and folic acid-modified polymer. The invention also discloses the preparation method and application of the aforementioned nano-preparation. The folic acid-modified nano-preparation containing the CCL19 recombinant expression gene can efficiently transfer the CCL19 recombinant expression gene into cells, improve the expression efficiency of CCL19, has excellent effect in treating colon cancer, and has good clinical application prospect.

Description

technical field [0001] The present invention relates to a nano preparation containing CCL19 recombinant expression gene and its preparation method and use. Background technique [0002] CCL19 (Chemokine C-C motif ligand 19) belongs to the CC class of chemokines, also known as Epstein-Barr virus-inducing molecule or human macrophage inflammatory protein 3β. It is mainly expressed in T cells in secondary lymphoid tissues and organs such as spleen and lymph nodes. naive T cells and mature DC cells. CCL19 plays a key role in anti-tumor, mainly through the following ways: induce dendritic cells to enter tumor tissue; mediate T cells involved in killing tumor cells; CCL19 expressed in tumor tissue induces immune cells to infiltrate tumors and release other cytokines; It directly binds to the CCR7 receptor and activates the post-receptor pathway to inhibit tumor proliferation, invasion and metastasis. CCL19 recruits immune cells to the tumor site, induces antigen presentation and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/51A61K47/34A61K47/18A61K47/22A61K48/00A61K38/19A61P35/00
CPCA61K9/5153A61K9/5123A61K48/005A61K38/195A61P35/00
Inventor 高祥王碧兰刘家刚程永忠
Owner SICHUAN UNIV