Application of GABRD methylation target in preparation of heroin relapse resisting drug

A technology of methylation and heroin, applied in the preparation of anti-heroin relapse drugs, in the field of methyltransferase inhibitors, can solve problems such as addiction, and achieve the effect of reducing relapse behavior

Inactive Publication Date: 2020-10-13
宁波市康宁医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term activation of opioid receptors by heroin leads to compensatory adaptation of opioid receptors and non-opioid receptor systems, leading to addiction

Method used

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  • Application of GABRD methylation target in preparation of heroin relapse resisting drug
  • Application of GABRD methylation target in preparation of heroin relapse resisting drug
  • Application of GABRD methylation target in preparation of heroin relapse resisting drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Establishment of heroin intravenous self-administration model

[0027] Intravenous Drug Self-Administration (Intravenous Drug Self-Administration) is one of the most classic animal models for studying drug addiction. Drug motivation and active compulsive drug use.

[0028] The classic rat heroin self-administration model can well simulate the heroin smoking behavior of clinical drug addicts. Rats can produce heroin self-administration after training. At the same time, the behavioral response rate and The dosage can reflect the compulsive drug-seeking behavior of drug addicts from the initial exposure to heroin to the addiction to heroin.

[0029] We used the following method to establish a self-administration model in rats: first, male SD rats underwent jugular vein cannulation, inserted a section of PE tube into the right jugular vein, and passed it out of the body through the back, antibacterial and recovery for more than a week after the operation . Subs...

Embodiment 2

[0030] Example 2 Illumina HiSeq sequencing

[0031] 12 hours after the heroin self-administration model was established stably, the rats were intraperitoneally administered with sodium pentobarbital, and sacrificed after deep anesthesia to isolate the NAc brain region. A heroin self-administration group (Heroin self-administration), a heroin passive administration group (Yoke heroin group) and a passive saline control group (Control) were established (n=3 for each group). The genomic DNA of the tissue samples was extracted with a PerkinElmer automatic nucleic acid extraction instrument (PerkinElmer Medical Diagnostic Products Shanghai Co., Ltd.), and the concentration of the obtained DNA was detected by a Qubit4.0 nucleic acid and protein analyzer. After DNA fragmentation and purification, end-repair and sequencing adapters were added, followed by Bisulfite treatment, library construction and quality inspection after amplification, and then Illumina HiSeq genome-wide methylation...

Embodiment 3

[0038] Example 3 Enlarging samples to detect changes in the DNA methylation level of the GABRD gene

[0039] According to Example 1, the rat heroin self-administration model was re-established, and a heroin self-administration group (Heroin self-administration), a heroin passive administration group (Yoke heroin group) and a passive saline control group (Control) were established. The NAc brain tissues of each group were collected, and the tissue DNA was extracted. The DNA methylation levels of 5 CpG sites in the DMR of the CpG island region of the GABRD gene were detected by bisulfite pyrosequencing technology. The basic principle of this technology: After the DNA sample is treated with bisulfite in the kit, the polymerase chain reaction (PCR) is used to amplify with methylation-specific amplification upstream and downstream primers, and then through specific sequencing primers Pyrosequencing was performed on the amplified products to calculate the degree of methylation at e...

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Abstract

The invention relates to application of a GABRD methylation target in preparation of a heroin relapse resisting drug. Compared with the prior art, the application has the advantages that the DNA methyltransferase inhibitor of the invention can reduce the methylation level of a GABRD gene and effectively reduce heroin relapse behaviors, indicating that the methyltransferase inhibitor targeting GABRD methylation can be used for treating heroin addiction and relapse.

Description

technical field [0001] The invention relates to a GABRD methylation target, in particular to the application of a methyltransferase inhibitor targeting GABRD methylation in the preparation of anti-heroin relapse drugs. Background technique [0002] Opioid addiction, especially heroin addiction, causes enormous medical and economic harm. Survey data from the United Nations Office on Drugs and Crime (UNODC) shows that globally, in 2018, 109,600 drug-related crimes were cracked, 137,400 criminal suspects were arrested, and 67.9 tons of various drugs were seized. Taking a relatively large proportion, the situation of heroin abuse is severe. Long-term activation of opioid receptors by heroin leads to compensatory adaptation of opioid receptors and non-opioid receptor systems, leading to addiction. Long-term chronic use of heroin leads to strong physical and mental dependence, which leads to a strong desire for drug use again, which is the main reason for the high relapse rate. ...

Claims

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Application Information

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IPC IPC(8): A61K31/706A61P25/36
CPCC12Q1/6858C12Q1/6883C12Q2600/136C12Q2600/154C12Q2537/164C12Q2531/113
Inventor 刘惠芬洪青晓徐文锦刘婧陈为升朱华强赖苗军周文华
Owner 宁波市康宁医院
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