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Method for using 5-flucytosine for real-time monitoring of deoxyribonucleic acid (DNA) deamination process of human immunodeficiency virus-1 (HIV-1)

A technology of HIV-1 and flucytosine, which is applied in the field of monitoring DNA deamination process, can solve the problems of signal overlap, inconvenient monitoring, difficult monitoring, etc., and achieve the effect of inhibiting HIV-1 virus infection

Inactive Publication Date: 2012-07-11
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method for real-time monitoring of the deamination process of HIV-1 virus DNA by human cytosine protein using 5-fluorocytosine, so as to overcome the deamination process of human cytosine protein deamination virus The reaction process of gene cytosine is very fast, it is not easy to monitor, and the use of hydrogen spectrum monitoring has defects such as signal overlap and inconvenient monitoring, which provides a convenient means for the research of inhibiting HIV-1 virus infection

Method used

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  • Method for using 5-flucytosine for real-time monitoring of deoxyribonucleic acid (DNA) deamination process of human immunodeficiency virus-1 (HIV-1)
  • Method for using 5-flucytosine for real-time monitoring of deoxyribonucleic acid (DNA) deamination process of human immunodeficiency virus-1 (HIV-1)
  • Method for using 5-flucytosine for real-time monitoring of deoxyribonucleic acid (DNA) deamination process of human immunodeficiency virus-1 (HIV-1)

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Embodiment

[0023] 1. Preparation of 4-(2,4,6-trimethylphenoxy)-5-fluorouracil derivatives:

[0024]

[0025] 1) Preparation of Compound 1

[0026] Dissolve 246mg of 5-fluoro-2'-deoxyuracil (1mmol) in dichloromethane, add 12mg of 4-dimethylaminopyridine (0.1mmol) and 0.18ml of pyridine (2.2mmol ), 0.3 ml of acetic anhydride (2.2 mmol). The reaction solution was stirred at room temperature for 12 h, and then 1 ml of methanol was added to continue stirring for half an hour to quench the reaction. The dichloromethane solvent was distilled off under reduced pressure, and 317 mg of a white foamy substance was obtained by silica gel column separation, with a yield of 96.1%.

[0027] m.p.143°C; Rf=0.52 (ethyl acetate:petroleum ether=3:1).

[0028] 1 H NMR (400MHz, CDCl 3 ): δH 8.78(d, J=3.6Hz, 1H), 7.655(d, J=6Hz, 1H), 6.281-6.319(m, 1H), 5.208-5.237(m, 1H), 4.269-4.433(m, 3H), 2.508-2.564(m, 1H), 2.17(dd, J 1’2’ =J 2’3’ =8Hz, 1H), 2.148(s, 3H), 2.122(s, 3H);

[0029] 13 C NMR (100M...

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Abstract

The invention firstly discloses a method for using 5-flucytosine for real-time monitoring of deoxyribonucleic acid (DNA) deamination process of human immunodeficiency virus-1 (HIV-1). Firstly, a 5-flucytosine precursor is led in a HIV-1 single-stranded DNA sequence through a solid-phase synthesis method to compound a DAN sequence containing the 5-flucytosine; then the synthetic DNA sequence is mixed with a humanized cytosine protein, and the DNA deamination reaction process is monitored through variation of F spectrum signals. The method solves the problem that a reaction process of humanized cytosine protein deamination virus gene cytosine is very fast and not easy to monitor, signal overlapping and monitoring inconvenience exist when hydrogen spectrum monitoring is used and the like, and provides a convenient and fast method for research of inhibition of HIV-1 virus infection.

Description

technical field [0001] The invention relates to a method for monitoring the deamination process of DNA, in particular to a method for real-time monitoring of the deamination process of HIV-1 virus DNA by human cytosine protein using 5-fluorocytosine. Background technique [0002] Studies have shown that: HIV-1 virus infects the human body through its cDNA replication in the human body, and the human body uses its own immune function to protect the body from infection; the DNA cytosine deamination protein APOBEC3G of the human immune system, which is expressed in the spleen, testis, and ovary , blood leukocytes such as T-lymphocytes and macrophages are abundantly expressed. This protein can inhibit the HIV-1 virus infection of the virus infection factor Vif protein deficiency. The specific mechanism is: APOBEC3G protein can specifically deaminate HIV-1 1 The two cytosines at the 3' end of the smallest cDNA fragment 5'-CCC-3' just synthesized in the human body are changed to u...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N7/01C12N15/09C12R1/93
Inventor 曹春阳姚祝军魏建科刘珊珊蓝文贤
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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