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Antitumor genetic transcription regulating medicine

A technology of transcriptional regulation and medicine, which is applied in the direction of antineoplastic drugs, gene therapy, drug combination, etc., to achieve the effect of significant drug effect, improved drug effect, and less dosage

Inactive Publication Date: 2004-06-23
JIANGSU KEYGEN BIOTECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research on decoy nucleic acid is limited to a few transcription factors, such as: CREB, NFkaapaB, E2F, etc.

Method used

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  • Antitumor genetic transcription regulating medicine
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation and analysis of K200 series decoy nucleic acid drugs

[0024] The 20 K200 series decoy nucleic acid drug sequences are:

[0025] Drug K201: 5'-CGCCCGCCGG CG CCCCAGGC GCCGGCGGGC G-3' (SEQ ID NO: 6) single chain.

[0026] Drug K202: 5'-CGCCCGCGGCCGGCCGCGGG CG-3' (SEQ ID NO: 7) single chain.

[0027] Drug K203 5'-CTGACCGCCTGAGGCGGTCAG-3' (SEQ ID NO: 8) single chain.

[0028] Drug K204: 5'-CTCGCCTGAGGCGTTAC-3' (SEQ ID NO: 9) double strand.

[0029] Drug K205: 5'-CCTGTAGCCTGAGGCTACAGG-3' (SEQ ID NO: 10) single chain.

[0030] Drug K206: 5'-CATCGCCCCGGGGCATC-3' (SEQ ID NO: 11) double strand.

[0031] Drug K207: 5'-ACTGACCGCCCGCGGCCCGT-3' (SEQ ID NO: 12) double strand.

[0032] Drug K208: 5'-CGCCCCGGGGCGGCCGCCCCGG-3' (SEQ ID NO: 13) single strand,

[0033] Drug K209: 5'-ACGCCGGGGGCGGGGTCA-3' (SEQ ID NO: 14) double strand.

[0034] Drug K210: 5'-GTGCCAGGGGCGGCCCCTGGCAC-3' (SEQ ID NO: 15) single chain.

[0035] Drug K211: 5'-CCCCGCCCCTGAGGGGCGGGG-3'...

Embodiment 2

[0046] Example 2 The growth inhibition of 20 decoy nucleic acids of K200 series on lung cancer A549 cells and liver cancer cells SMMC7721

[0047] Use RMPI1640 culture solution containing 10% newborn bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin at 37°C and 5% CO 2 The cells in good growth state were digested with trypsin, counted, inoculated in a 96-well cell culture plate, the number of cells per well was 10,000, and the cells were placed at 37°C, 5% CO 2 cultured overnight in an incubator, in the absence of serum, using LipofectANINE TM , and refer to the kit instructions for decoy oligonucleotide transfection, the concentration of decoy oligonucleotide is 200nmol / L, each concentration is 4 wells, the total volume of each well is 100μL, and the cell control well is set, the cell control Add the same concentration of LipofectANINE into the well TM RMPI1640 culture medium. After 5 hours of transfection, the culture medium was changed to RMPI1640 medium cont...

Embodiment 3

[0048] Example 3 In vivo anti-tumor efficacy test of decoy nucleic acid drug K201 in animals

[0049] The decoy nucleic acid drug K201 was prepared with normal saline to an appropriate concentration, and stored in a refrigerator at 4°C until use.

[0050] T cell immunodeficiency nude mice (18-22g, 6 weeks, female, placed in a laminar flow frame and raised under SPF conditions, free to ingest food and water. The tumor cells cultured in vitro were inoculated into the third generation of nude mice grown in vivo The tumor was used as the tumor source for the test, and the vigorously growing tumor source was taken under sterile conditions, and prepared into about 1×10 7 / ml cell suspension, subcutaneously inoculate 0.2ml / every mouse in nude mice, after inoculation 24 hours, nude mice are divided into four groups, the daily intravenous injection medicine 0.1ml of administration group, dosage is respectively 1.0mg / kg, 5.0 mg / kg, 10.0mg / kg, the control group is the corresponding norma...

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Abstract

The present invention belongs to the field of genetic treating medicine of tumor, and is antitumor genetic transcription regulatory medicine. The medicine component consists of 15-40 nucleotides, contains decoy nucleic acid with specific sequence 5í»-SCCNNNVSS-3í», may be combined specifically with transcription factor AP-2 via competition with several genes containing cis element with the factor combining site, and can regulate gene expression and suppress malignant amplification of tumor to treat tumors.

Description

Field of invention: [0001] Tumor Gene Therapy Drugs Background technique: [0002] Gene therapy for tumors is a specific and advanced treatment method at the molecular level. However, due to the large size of the gene molecule, it is difficult to enter the cell, the operation of the virus vector is complicated, and the implementation is difficult. There are still many problems to be solved before the actual application. A new method of regulation to realize tumor gene therapy. Decoy nucleic acid is an artificially designed and synthesized double-stranded oligonucleotide. The molecule is much smaller than the gene. It has a high affinity with the target transcription factor and can enter the cell as a decoy cis element. By inhibiting specific transcription factors and The combination of regulatory regions can change the expression of genes to achieve the purpose of treating tumors. But the current research on decoy nucleic acid is limited to a few transcription factors, suc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7088A61K48/00A61P35/00
Inventor 叶青王雪根陈武领冯莹刘晖韦策
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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