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Compound containing anti-PD (Program Death)-1 gene and poly-spermidine and application of compound to treatment of tumors

A technology of polyspermidine and complexes, which is applied in the field of Ming Dynasty, can solve problems such as affecting normal distribution, and achieve the effect of improving the transfection effect.

Inactive Publication Date: 2017-02-15
SHANGHAI YUNSHUN BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cationic liposomes have high transfection activity in vivo and in vitro, however, because the positive charge on the surface affects its normal distribution in the body, and at the same time, due to the selection of cationic lipids, immunogenicity and inflammatory reactions have become inevitable in animal experiments One of the shortcoming (Gao, K.&Huang, L.Nonviral methods for siRNA delivery. Molecular pharmaceuticals 6,651-658(2008).)

Method used

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  • Compound containing anti-PD (Program Death)-1 gene and poly-spermidine and application of compound to treatment of tumors
  • Compound containing anti-PD (Program Death)-1 gene and poly-spermidine and application of compound to treatment of tumors
  • Compound containing anti-PD (Program Death)-1 gene and poly-spermidine and application of compound to treatment of tumors

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preparation example Construction

[0033] The preparation method of the compound containing anti-PD-1 gene and polyspermidine includes the following steps:

[0034] The aqueous solution of the polyspermidine polymer and the aqueous solution of the anti-PD-1 gene are mixed, and the complex is prepared by standing at room temperature for 20-60 minutes.

[0035] Application of the compound containing anti-PD-1 gene and polyspermidine in the treatment of tumors.

[0036] The method for treating tumors using a complex containing anti-PD-1 gene and polyspermidine includes the following steps:

[0037] The complex containing the anti-PD-1 gene and polyspermidine is co-cultured with T cells to prepare T cells that secrete anti-PD-1 antibodies, and then inject T cells that secrete anti-PD-1 antibodies. Just enter the tumor patient.

Embodiment 1

[0038] Example 1 Preparation of complex formed by polyspermidine polymer (PSAi) and anti-PD-1 plasmid DNA

[0039] The polyspermidine polymer prepared by polyspermidine and the linking agent 2,6-pyridinediformaldehyde is formulated into an aqueous solution with a concentration of 2μg / μl and diluted with sterile water to 20ng / μl, 40ng / μl, 100ng / μl , 200ng / μl, 1000ng / μl and 2000ng / μl solutions. In addition, the anti-PD-1 gene plasmid DNA was made into an aqueous solution with a concentration of 20ng / μl. Add an equal volume of the polyspermidine polymer aqueous solution to the plasmid DNA aqueous solution, vortex for 5 minutes, and let stand for 20 minutes to obtain a series of nanocomposite solutions.

Embodiment 2

[0040] Example 2 The particle size and potential of the complex formed by polyspermidine polymer (PSAi) and anti-PD-1 plasmid DNA Characterization

[0041] Put the series of complex solutions prepared in Example 1 into the test tank of the BIC90plus particle size potential analyzer. The mass ratios of PSAi and plasmid DNA in the complexes of the series of complex solutions are 0.25, 0.5, 2, 5, 10. 50, the test temperature is set to 25°C, the medium is water, the viscosity is 0.89cp, the refractive index is 1.330, and the light scattering angle is 90. , The detection wavelength is 659nm, each sample is tested 3 times, each run time is 2min, and the average particle size is recorded. The result is figure 1 As shown, the results show that the particle size of the nanocomposite is below 200 nm. Its dispersion index meets the requirements.

[0042] Put the series of complex solutions prepared in Example 1 into the test tank of the BIC90plus particle size potential analyzer. The mass...

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Abstract

The invention relates to the technical field of T cell tumor treatment and in particular relates to a compound containing an anti-PD (Program Death)-1 gene and poly-spermidine and application of the compound to treatment of tumors. The compound contains a polyspermidine polymer and the anti-PD-1 gene at the mass ratio of (1 to 100) : 1. The compound is transfected to a T cell in vitro to prepare a self-secretion anti-PD-1 antibody-T cell and can generate a PD-1 antibody, so as to help to block a PD-1 signal and effectively help a depleted T cell, and promote the T cell capable of resisting tumor immune response to treat the tumors. Compared with the prior art, the secretion anti-PD-1 antibody-T cell provided by the invention can be used for effectively treating the tumors, and immune treatment of the tumors can be realized; an in-vivo toxicity problem does not exist.

Description

Technical field [0001] The present invention relates to the technical field of tumor treatment, in particular to a compound containing anti-PD-1 gene and polyspermidine and its application in the treatment of tumors, in particular to a kind of anti-PD-1 (programmed death-1 ) The nanocomplex formed by gene and polyspermidine is transfected into T cells in vitro to prepare T cells that secrete anti-PD-1 antibodies, which can produce PD-1 antibodies to help block PD-1 signals. A method of T cell treatment of tumors that effectively helps depleted T cells and promotes anti-tumor immune response. Background technique [0002] T lymphocytes target tumor cells that express this specific antigen. Because of its characteristics of targeted killing of tumor cells, CTL therapy has become an ideal solution for individualized tumor treatment, and has attracted more and more attention from immunologists in the world. However, due to the production of PD-1, T cells lose their activity. In or...

Claims

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Application Information

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IPC IPC(8): C12N15/87A61K35/17A61P35/00
CPCA61K35/17C12N15/87
Inventor 马孟
Owner SHANGHAI YUNSHUN BIOTECH
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