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Nucleic acid hydrolysis cutting agent based on cucurbituril

A cutting agent, nucleic acid technology, used in gene therapy, sugar derivatives, organic chemistry, etc., can solve problems such as hindering applications

Inactive Publication Date: 2007-03-28
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are relatively few artificial enzyme systems that can effectively catalyze the hydrolysis of DNA
Moreover, the vast majority are complexes of metal ions. Most of these chemical nucleases containing metal ions require an acidic optimum pH and the activity of these enzymes is controlled by metal ions, which hinders them in chelation. chelating buffer or in the presence of metal chelating agents

Method used

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  • Nucleic acid hydrolysis cutting agent based on cucurbituril
  • Nucleic acid hydrolysis cutting agent based on cucurbituril
  • Nucleic acid hydrolysis cutting agent based on cucurbituril

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Step 1. Synthesis of the main body cucurbit ring molecule:

[0045] 11.4g glycoluril (80mmol) in 100ml four-neck round bottom flask, add 40ml concentration and be the sulfuric acid H2SO4 solution of 9M (mol / liter), oil bath is warming up to 70 ℃, begins to drop 14ml, 37% formaldehyde aqueous solution, drops When adding, keep the temperature between 70-75°C. After the dropwise addition, keep the temperature at 70-75°C for 24 hours, then raise the temperature to 95-100°C, continue the reaction for 12 hours, cool to room temperature, and pour the reaction solution into 400ml of water , 2 L of acetone was added, and a white solid was precipitated. After standing for 20 min, the obtained solid was collected by suction filtration. The obtained solid was dispersed in 200°C of water, added with 200ml of acetone, stirred for 20min, left to stand, and the white solid obtained by suction filtration was washed three times with 300ml of water at 60°C, rinsed with about 50ml of aceto...

Embodiment 2

[0051] Step 1. Repeat step 1 of Example 1 to obtain the main body melon ring used.

[0052] Step 2. Use concentrated phosphoric acid to replace the concentrated hydrochloric acid used in Step 2 of Example 1 to obtain the guest molecule G2, 1,6-diimidazolylhexane diphosphate. Elemental analysis (calcd.%) for C 12 h 24 N 4 P 2 o 8 : C, 34.78; N, 13.53; H, 5.80. Found: C, 34.91; N, 13.74; H, 5.73.

[0053] Step 3. Substitute the guest molecule G1 with the guest molecule G2, and repeat the step 3 of Example 1 to obtain the quasi-rotaxane compound P2 with a yield of 86%. Elemental analysis (calcd.%) for C 36 h 36 N 24 o 12 ·C 12 h 20 N 4 2H 2 PO 4 12H 2 O: C, 35.42; N, 24.11; H, 5.17. Found: C, 35.59; N, 24.32; H, 5.07.

Embodiment 3

[0055] Step 1. Repeat step 1 of Example 1 to obtain the main body melon ring used.

[0056] Step 2. The concentrated hydrochloric acid used in Step 2 of Example 1 was replaced with concentrated sulfuric acid to obtain the guest molecule G3, 1,6-diimidazolylhexane disulfate. Elemental analysis (calcd.%) for C 12 h 22 N 4 8 2 o 8 : C, 34.78; N, 13.53; H, 5.31.Found: C, 34.78; N, 13.64; H, 5.22.

[0057] Step 3. The guest molecule G1 was replaced by the guest molecule G3, and step 3 of Example 1 was repeated to obtain the quasi-rotaxane compound P2 with a yield of 84%. Elemental analysis (calcd.%) for C 36 h 36 N 24 o 12 ·C 12 h 20 N 4 ·2HSO 4 10H 2O: C, 36.23; N, 24.65; H, 4.91. Found: C, 36.39; N, 24.72; H, 4.88.

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Abstract

The invention relates to pseudorotaxane molecular compound produced by cucurbit[6]uril and 1, 6-di-imidazole group di-acid salt whose chemical molecular formula is C36H36N24O12*C12H20N4*2X*nH2O whose X equals to Cl, H2PO4, HSO4, NO3, Br; n is not less than 1. The compound can be used in nucleic acid cutting whose condition is that the cutting reagent is dissolved in buffer system solution with given pH value, then mixed with cutting substrate; their final density is kept in given range; and their temperature is also kept in given range. The cutting reagent can be used to cut nucleic acid in short time at physiology condition, further study the application in gene therapy medicine preparation to develop into gene therapy medicine.

Description

technical field [0001] The invention relates to a nucleic acid cutting agent, in particular to a cucurbit ring-based nucleic acid hydrolysis cutting agent. Background technique [0002] As the carrier of genetic information and the basis of gene expression, accounting plays a very important role in the growth, development, reproduction and other life activities of organisms. The completion of HGP in 2000 marks that human medicine will enter a gene and molecular era. The secrets of life, genetic diseases, molecular diseases, cancer, AIDS, blood supply, and organ supply that have plagued human beings for thousands of years can find scientific interpretations. and a way out. The core of all this is the recognition, tailoring, and connection of DNA nucleic acid molecules. Therefore, the research on artificial nucleic acid cleavage reagents has always been the most active research topic in biochemistry and molecular biology. Nucleic acid cleavage is the breaking of long-chain ...

Claims

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Application Information

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IPC IPC(8): C07H21/04A61K48/00C07D233/61C07D487/22
Inventor 霍方俊阴彩霞杨频
Owner SHANXI UNIV
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