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Composite for treating cancer containing oligonucleotide and nontoxic LPS

A technology of composition and deoxynucleotide, which can be used in medical preparations containing active ingredients, drug combinations, organic active ingredients, etc., and can solve problems such as sepsis

Inactive Publication Date: 2009-03-25
DAEWOONG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the problem is that, as a typical endotoxin, LPS is highly toxic
In addition, the binding of ordinary LPS to DNA can cause severe symptoms such as sepsis

Method used

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  • Composite for treating cancer containing oligonucleotide and nontoxic LPS
  • Composite for treating cancer containing oligonucleotide and nontoxic LPS
  • Composite for treating cancer containing oligonucleotide and nontoxic LPS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: screening avirulent strain

[0037] Screening and discovery of Escherichia coli mutants with very short lipopolysaccharide

[0038] The Escherichia coli (E.coli) strain EG0021 having a very short lipopolysaccharide sugar chain was discovered from Escherichia coli living in the intestinal tract of healthy people, and the Escherichia coli (E.coli) strain EG0021 was established Method for purifying lipopolysaccharide.

[0039] A single colony of Escherichia coli (E. coli) obtained from a healthy male adult was cultured in a liquid medium, followed by repeating the selection operation 5 times to obtain 50 strains of Escherichia coli (E. coli). And, get each bacterium colony from the 50 strains selected on the plate, dissolve in 4ml of 0.9% physiological saline, then take 1ml of the resulting solution and transfer it to an Eppendorf centrifuge tube (Eppendorf tube), and use 2 μl DNase I treatment was performed, followed by reaction at 37° C. for 1 hour in...

Embodiment 2

[0041] Example 2: CG Methylation

[0042] To characterize the function of unmethylated CGs in oligonucleotides, cytosine residues of CG sequences were selectively methylated using SssI methylase.

[0043] DNA methylation was performed by mixing 1 unit of CpG methylase (M.Sss I; NEB M0226S) with 10 µg of ODN7909, followed by mutual reaction at 37°C for 12 hours. At this time, 160 µ of MS-adenosylmethionine (SAM) as a methyl donor was mixed and reacted together.

[0044] After complete methylation, residual salts and enzymes were then removed using DNA cleanup kit (CPG DPC60050) and Micropure-EZ (Amicon 42529).

Embodiment 3

[0045] Example 3: Purification of CIA02 from mutant Escherichia coli (E.coli)

[0046] Purification of lipopolysaccharide from mutant Escherichia coli (E.coli)

[0047] Escherichia coli (E. coli) strains were prepared in the same manner as in DNA isolation.

[0048] The strain was then mixed with 2 volumes of ethanol, centrifuged at 4,000 g to settle the pellet, and then 1.5 volumes of acetone was added to the resulting pellet, thoroughly mixed and centrifuged at 4,000 g.

[0049] An equal amount of diethyl ether was added to the resulting precipitate, mixed thoroughly and centrifuged at 4,000 g. The cell pellet obtained by centrifugation was covered with perforated aluminum foil, dried, and the cell body was weighed, followed by adding an extraction mixture (90% phenol:chloroform:petroleum ether=2:5: 8).

[0050] The resulting mixture was divided into glass centrifuge tubes, and centrifuged at 25° C. and 3,000 rpm (1,200 g) for 20 minutes to obtain a supernatant. The ...

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Abstract

Disclosed is a composition for treating cancer including oligodeoxynucleotides and LPS-derived non-toxic lipopolysaccharides as effective components.

Description

technical field [0001] The present invention relates to an anticancer drug using a non-toxic high molecular compound (CIA05) and oligodeoxynucleotide (ODN) derived from LPS. Background technique [0002] Methods of treating cancer that have evolved since the 1960s are mainly divided into three groups: surgery, radiation therapy, and chemotherapy. In the United States, the fact that cancer mortality rates had been rising, and had risen in the past, until 1973, when cancer mortality rates suddenly decreased, speaks to the success of the treatment approach. However, surgery and immunotherapy have limitations, namely since they are limited to local treatments, they have a good prognosis if cancer in situ is disturbed at an early stage. Also, chemotherapy was successful in killing all cancer cells. In this case, elderly and debilitated patients may lose their lives because the host, the normal tissues of the patient, especially the immune tissues, are also severely damaged. ...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61P35/00A61K31/739
CPCA61K31/739A61K31/7088A61P35/00Y02A50/30A61K2300/00
Inventor 安宝暎赵良济俞元一李奈暻金斗植
Owner DAEWOONG CO LTD
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