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Anti-sense nucleotide of c-erbB2 cancer gene combined with epidermal growth factor and its application

A technology of epidermal growth factor and c-erbb2, applied in gene therapy, sugar derivatives, drug combinations, etc., can solve problems affecting tumor diagnosis and treatment effects, and small target/non-target ratio

Inactive Publication Date: 2007-09-19
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In antisense gene research, the most important thing is how many antisense short ribonucleotides (ASON) enter the cell and combine with the target sequence to play an antisense inhibitory effect on tumors. This is radionuclide antisense gene imaging. And the key to the success of antisense gene therapy. Under the state of the art, after the antisense gene nucleotides with c-erbB2 as the target gene enter the cells, the target / non-target ratio is small, which affects the diagnosis and treatment of tumors

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  • Anti-sense nucleotide of c-erbB2 cancer gene combined with epidermal growth factor and its application

Examples

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Embodiment 1

[0036] Example 1 The preparation method of c-erbB2 oncogene antisense short nucleotide combined with epidermal growth factor includes the following steps:

[0037] (1) Synthesis of EGF-polylysine conjugate (EGF-PL)

[0038] Put 500μg EGF, 2.5mg polylysine (PL), 2.6mg EDAC and 50μl tris-hydrochloric acid buffer (0.1M / L, pH=7.3) in the reaction flask, and react under stirring at room temperature 30h, carefully put the reactant into a dialysis bag, and dialyze it in HBS buffer (150mM / L NaCl, 20mM / L Hepes acid, pH=7.3) for 25h, and change the dialysate twice during this period to obtain EGF-PL HBS Solution (concentration is 5u g / μl);

[0039] (2) EGF-PL forms a complex with antisense short ribonucleotides (ASON)

[0040] The EGF-PL and ASON eluate were mixed at a molar ratio of 20:1 and reacted at room temperature for 25 minutes. The mixture was analyzed by agarose / tris-acetic acid buffer (1×) electrophoresis (100V, 30min) to confirm the formation EGF-PL-ASON complex.

Embodiment 2

[0041] Example 2 Imaging Research

[0042] Imaging of tumor-bearing nude mice, EGF- 99m One hour after Tc-ASON was injected into tumor-bearing nude mice, the tumor tissue began to concentrate radioactively. The main distribution in the body was liver and kidney. The radioactivity in the tumor tissue was high. Injecting 0.1mCi can ensure successful imaging.

[0043] The tumor was clearly imaged in 2 hours, the tumor image became lighter in 3 hours, and gradually faded in 4 hours. Using this performance can provide relevant information for the shape, size and number of tumors.

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Abstract

The invention provides a antisense short nucleotide of c-erbB2 cancer gene whose sequence is: SEQ ID NO 1. The antisense short nucleotide is combined with epidermal growth factor by Poly-L-Lysine, injecting the vinculum into tumour cell of body, EGFR receptor and c-erbB2 both belong to epidermal growth factor receptor family with distributed conformity and consanguinity in organisation, so increasing ratio of target / nontarget, raising diagnostic sensibility and treatment effect of antisense gene for excess expressed cancer of c-erbB2 cancer gene.

Description

Technical field [0001] The invention relates to an antisense short nucleotide, in particular to an EGFR-mediated c-erbB2 oncogene antisense short nucleotide that enhances tumor targeting and its application. Background technique [0002] With the development of medical biotechnology, humans have entered the molecular level in the diagnosis and treatment of malignant tumors, and antisense genes have been used in the field of targeted diagnosis and targeted therapy of tumors. An antisense gene refers to a nucleotide sequence that is complementary to a target gene and can inhibit gene expression. The antisense gene technology uses oncogene DNA or ribonucleic acid mRNA as a target sequence, artificially synthesized complementary to the target sequence The 13-20mer single-stranded antisense short ribonucleotide (ASON) can specifically bind to the target sequence to block gene expression and inhibit tumor growth. Or label the antisense short ribonucleotides with radionuclides to make a...

Claims

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Application Information

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IPC IPC(8): C12N15/11C07H21/04A61K31/7088A61K48/00A61P35/00C12Q1/68C12N15/10C12N15/113
Inventor 李少林吴永中季平段红王颖彭志平杜铭庞华谢娟黄家君李兵傣家富刘方欣
Owner CHONGQING MEDICAL UNIVERSITY
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