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Tumor blood vessel-specific chemotaxis CTL nanosystem and its preparation and application

A nanoparticle and nanoliposome technology, which is applied in the direction of antineoplastic drugs, medical preparations containing active ingredients, peptide/protein components, etc., to achieve the effect of reducing growth rate and prolonging survival time

Inactive Publication Date: 2018-08-07
GUANGXI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, tumors are mainly enriched with IP-10 through transgenic technology or direct local multi-point injection of tumors. The former has targeting problems, while the latter can only increase the concentration at the injection site.

Method used

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  • Tumor blood vessel-specific chemotaxis CTL nanosystem and its preparation and application
  • Tumor blood vessel-specific chemotaxis CTL nanosystem and its preparation and application
  • Tumor blood vessel-specific chemotaxis CTL nanosystem and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Embodiment 1, the preparation of the nanoliposome (ENG-Apt / IP-10-LP) of the ENG-Aptamer modification of load IP-10 gene

[0090] The gene delivery system provided by the present invention is a nanoparticle composed of a core and a shell, which is an ENG-Apt-modified nanoliposome (ENG-Apt / IP-10-LP) loaded with an IP-10 gene;

[0091] The shell is composed of aptamers (Endoglin-Aptamer, ENG-Apt), cross-linking agents and nanoliposomes;

[0092] The sequence of the aptamer is shown in SEQ ID No.1; the aptamer is connected to the nanoliposome through a cross-linking agent; the cross-linking agent is DSPE-PEG 2000 -Mal;

[0093] The core is the vector GV143 IP-10 containing the IP-10 gene, GV143 IP-10 expresses IP-10 shown in SEQ ID No.2, and the nucleotide sequence of the IP-10 gene in GV143 IP-10 is SEQ ID No.2 in the sequence table ID No.3, encoding IP-10 shown in SEQ ID No.2.

[0094] 1. Preparation of ENG-Apt / IP-10-LP

[0095] Preparation of S1, Nanoliposome (LP)

...

Embodiment 2

[0108] Example 2, the targeting effect of ENG-Aptamer modified nanoliposomes loaded with IP-10 gene on tumor cells

[0109] The ENG-Apt / IP-10-LP of Example 1 were labeled with green fluorescent FITC (fluorescein isothiocyanate) and red fluorescent DiI respectively to obtain FITC-labeled ENG-Apt / IP-10-LP and DiI-labeled Labeled ENG-Apt / IP-10-LP; FITC-labeled ENG-Apt / IP-10-LP and DiI-labeled ENG-Apt / IP-10-LP were incubated with 293T-mE at 37°C for 45min , observed under a microscope, the results are as follows Figure 4 Shown in A. In addition, 293T-mE was replaced by 293T cells, and other steps remained unchanged as a control.

[0110] Incubate non-fluorescence-labeled ENG-Aptamer with 293T-mE for 30 min in advance, then add FITC-labeled ENG-Apt / IP-10-LP and DiI-labeled ENG-Apt / IP-10-LP respectively, at 37°C Incubate for 45min, observe under the microscope, the results are as follows: Figure 4 Shown in A. In addition, 293T-mE was replaced by 293T cells, and other steps rema...

Embodiment 3

[0113] Embodiment 3, the therapeutic effect of nanoliposomes modified by ENG-Aptamer loaded with IP-10 gene on tumor

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Abstract

The invention discloses a tumor vessel-specific chemotactic CTL nanometer system and its preparation and application. The system provided by the present invention is a nanoparticle composed of a core and a shell; the shell is composed of an aptamer, a cross-linking agent and a nanoliposome; the sequence of the aptamer is shown in SEQ ID No.1 The aptamer is connected with the nanoliposome by the cross-linking agent; the core is IP-10, and the IP-10 is the protein of the following B1) or B2): B1) the amino acid sequence is The protein of SEQ ID No.2; B2) derived from B1) with the same function through substitution and / or deletion and / or addition of one or several amino acid residues in the amino acid sequence shown in SEQ ID No.2 protein. Experiments have proved that the gene delivery system of the invention can be used to treat tumors or prepare anti-tumor drugs or vaccines.

Description

technical field [0001] The invention relates to a tumor blood vessel-specific chemotactic CTL nanometer system in the field of biomedicine and its preparation and application. Background technique [0002] Tumor is one of the major diseases that seriously endanger human health. Tumor antigen-specific cytotoxic T lymphocyte (cytotoxic T lymphocyte, CTL) adoptive therapy has been one of the research hotspots and difficulties in tumor immunotherapy. Years of studies on tumor-specific CTL adoptive therapy often achieve good results in cell experiments, but the animal experiments or clinical effects are not good. The important reason is that the number of tumor-specific CTL cells that can target the tumor site in the body is limited, and due to the immune escape mechanism, the biological activity of specific CTL targeting the tumor site to recognize and kill tumor cells is greatly inhibited, making it difficult to Efficient and thorough removal of tumor cells. Therefore, in or...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K38/19A61K9/127A61K35/74A61K36/06A61K35/12A61K35/00A61K36/00A61P35/00
Inventor 卢小玲赵永祥
Owner GUANGXI MEDICAL UNIVERSITY