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Nano gene drug capable of stimulating lymphocyte activation and preparation method thereof

A nano and gene technology, applied in gene therapy, drug combination, pharmaceutical formulation, etc., can solve the problems of inability to aggregate, low efficiency of transfecting cells, and unsatisfactory treatment effects of tumors or infectious diseases, etc. Proliferating tumor targeting, enhancing drug efficacy, and prolonging half-life effects

Inactive Publication Date: 2015-09-09
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] When a simple DNA gene drug is applied in vivo, the active uptake efficiency of cells is low, the efficiency of transfection cells is not high, and the expression of related gene products in vivo is affected by the injection route and immune dose
In addition, the expression products of DNA gene drugs generally cannot accumulate in tumors or local infection sites in vivo, resulting in the therapeutic effect on tumors or infectious diseases often not reaching the desired effect

Method used

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  • Nano gene drug capable of stimulating lymphocyte activation and preparation method thereof
  • Nano gene drug capable of stimulating lymphocyte activation and preparation method thereof
  • Nano gene drug capable of stimulating lymphocyte activation and preparation method thereof

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Experimental program
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Effect test

Embodiment Construction

[0043] 1. Construction of pcDNA3.1-sig-dsNKG2D-IL-21 fusion gene eukaryotic expression vector:

[0044] 1.1 Primer sequences for PCR amplification: Using bioinformatics means, design corresponding primers (see Table 1), and amplify signal peptides-sNKG2D(1), sNKG2D(2), IL -21 gene segment. The reverse complementary sequence of (Gly4Ser)4 (shown in the box in the table) was added to the downstream primer of signal peptide-sNKG2D (1) and the downstream primer of sNKG2D (2). figure 2 The construction strategy of the ATG-sig-dsNKG2D-IL-21 fusion gene fragment is shown.

[0045] Table 1. Primer sequences for amplification of signal peptide, NKG2D, and IL-21 gene fragments

[0046]

[0047] The sequence of the PCR amplification product of the signal peptide-sNKG2D(1) is shown in SEQ ID NO: 4, the 1st-6th base is the sequence of the Xba I restriction site; Randomly added nucleotides; No. 8-68 is the gene sequence of mouse β2-microglobulin signal peptide; No. 69-485 is the gene...

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Abstract

The invention relates to a novel nano gene drug capable of stimulating lymphocyte activation and a preparation method thereof. The invention discloses a fusion gene; and a mouse beta-2 microglobulin signal peptide is introduced from the upstream of the fusion gene dsNKG2D-IL-21 gene to obtain the fusion gene ATG-sig-dsNKG2D-IL-21. The invention also discloses a fusion gene expression carrier pcDNA3.1-sig-dsNKg2D-IL-21. The invention also discloses a nano gene vaccine drug which is formed by encapsulating the pcDNA3.1-sig-dsNKg2D-IL-21 fusion protein expression vector with chitosan. The invention also discloses a preparation method of a nano gene vaccine. The nano drug is used for carrying a nano gel of the recombinant dsNKG2D-IL-21 gene vector capable of stimulating lymphocyte activation and in-vivo targeted distribution, and is hopeful to become a drug for resisting tumors and infectious diseases.

Description

technical field [0001] The invention relates to a novel nano-gene drug for stimulating lymphocyte activation and a preparation method thereof, belonging to the field of biotechnology. Background technique [0002] The activation of lymphocytes plays an important role in the body's anti-infection and anti-tumor functions. However, in the process of persistent chronic infection or tumorigenesis, the immune function of the body is suppressed and is in a low activity state. The preparation of effective drugs for stimulating lymphocyte activation is of great significance for the effective treatment of chronic infectious diseases and tumors. The growth and activation of lymphocytes mainly depend on cytokines, such as IL-2, IL-7, IL-15, IL-21, etc. Therefore, based on the genes and protein sequences of cytokines, the corresponding genes and proteins are prepared in vitro, theoretically It has a good function of promoting lymphocyte proliferation and stimulating lymphocyte activat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/63A61K48/00A61K39/00A61P35/00A61P37/02
Inventor 赵培蕾龚卫娟肖炜明
Owner YANGZHOU UNIV
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